Journal of extracellular biology最新文献

{"title":"Correction to A thermo-resistant and RNase-sensitive cargo from Giardia duodenalis extracellular vesicles modifies the behaviour of enterobacteria","authors":"","doi":"10.1002/jex2.114","DOIUrl":"https://doi.org/10.1002/jex2.114","url":null,"abstract":"<p>Siddiq, A., Dong, G., Balan, B., Harrison, L. G., Jex, A., Olivier, M., Allain, T., &amp; Buret, A. G. (2023). A thermo-resistant and RNase-sensitive cargo from <i>Giardia duodenalis</i> extracellular vesicles modifies the behaviour of enterobacteria. <i>Journal of Extracellular Biology</i>, 2, e109. https://doi.org/10.1002/jex2.109</p><p>In the originally-published article, incorrect text was included for the Acknowledgements and Conflict of Interest Statement sections. The correct text appears below. The article has been updated online.</p><p>We apologize for this error.</p><p>All co-authors have seen and agree with the contents of the manuscript and there is no conflict of interest</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-09-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.114","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50136332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"High resolution imaging and analysis of extracellular vesicles using mass spectral imaging and machine learning","authors":"Sarah Elizabeth Bamford,&nbsp;Natasha Vassileff,&nbsp;Jereme G. Spiers,&nbsp;Wil Gardner,&nbsp;David A. Winkler,&nbsp;Benjamin W. Muir,&nbsp;Andrew F. Hill,&nbsp;Paul J. Pigram","doi":"10.1002/jex2.110","DOIUrl":"https://doi.org/10.1002/jex2.110","url":null,"abstract":"<p>Extracellular vesicles (EVs) are potentially useful biomarkers for disease detection and monitoring. Development of a label-free technique for imaging and distinguishing small volumes of EVs from different cell types and cell states would be of great value. Here, we have designed a method to explore the chemical changes in EVs associated with neuroinflammation using Time-of-Flight Secondary Ion Mass spectrometry (ToF-SIMS) and machine learning (ML). Mass spectral imaging was able to identify and differentiate EVs released by microglia following lipopolysaccharide (LPS) stimulation compared to a control group. This process requires a much smaller sample size (1 µL) than other molecular analysis methods (up to 50 µL). Conspicuously, we saw a reduction in free cysteine thiols (a marker of cellular oxidative stress associated with neuroinflammation) in EVs from microglial cells treated with LPS, consistent with the reduced cellular free thiol levels measured experimentally. This validates the synergistic combination of ToF-SIMS and ML as a sensitive and valuable technique for collecting and analysing molecular data from EVs at high resolution.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.110","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50149802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Tetraspanin 15 depletion impairs extracellular vesicle docking at target neurons","authors":"Daniele Stajano,&nbsp;Franco L. Lombino,&nbsp;Michaela Schweizer,&nbsp;Markus Glatzel,&nbsp;Paul Saftig,&nbsp;Kira V. Gromova,&nbsp;Matthias Kneussel","doi":"10.1002/jex2.113","DOIUrl":"https://doi.org/10.1002/jex2.113","url":null,"abstract":"<p>Neurons in the central nervous system release extracellular vesicles (EVs) and exosomes in response to synaptic activity to regulate physiological processes at target neurons. The intercellular transfer of proteins, mRNAs, lipids or metabolites through EVs potentially modulates the structure and function of neurons and circuits. Whereas the biogenesis of EVs, their release from donor cells, and their molecular composition have been studied extensively, the critical factors and mechanisms regulating EV interactions with target cells are incompletely understood.</p><p>Here, we identified tetraspanin 15 (Tspan15) as a component of tumor susceptibility gene 101 protein (TSG101)- and CD81-positive EV fractions. Tspan15 fluorescent fusion proteins were released from donor cells and interacted with target cells together with the exosomal marker CD63. EVs collected from wildtype cortical neurons (WT-EVs) underwent similar association with target neurons derived from either wildtype (+/+) or Tspan15 knockout (−/−) mice. In contrast, target cell interactions of EVs collected from Tspan15 (−/−) cortical donor neurons (KO-EVs) were significantly impaired, as compared to WT-EVs. Our data suggest that Tspan15 is dispensable at target neuron plasma membranes, but is required at the EV surface to promote EV docking at target neurons.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.113","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50149801","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A thermo-resistant and RNase-sensitive cargo from Giardia duodenalis extracellular vesicles modifies the behaviour of enterobacteria","authors":"Affan Siddiq,&nbsp;George Dong,&nbsp;Balu Balan,&nbsp;Luke G. Harrison,&nbsp;Aaron Jex,&nbsp;Martin Olivier,&nbsp;Thibault Allain,&nbsp;Andre G. Buret","doi":"10.1002/jex2.109","DOIUrl":"10.1002/jex2.109","url":null,"abstract":"<p>Extracellular vesicles (EVs) recently emerged as important players in the pathophysiology of parasitic infections. While the protist parasite <i>Giardia duodenalis</i> can produce EVs, their role in giardiasis remains obscure. <i>Giardia</i> can disrupt gut microbiota biofilms and transform commensal bacteria into invasive pathobionts at sites devoid of colonizing trophozoites via unknown mechanisms. We hypothesized that <i>Giardia</i> EVs could modify gut bacterial behaviour via a novel mode of trans-kingdom communication. Our findings indicate that <i>Giardia</i> EVs exert bacteriostatic effects on <i>Escherichia coli</i> HB101 and <i>Enterobacter cloacae</i> TW1, increasing their swimming motility. <i>Giardia</i> EVs also decreased the biofilm-forming ability of <i>E. coli</i> HB101 but not by <i>E. cloacae</i> TW1, supporting the hypothesis that these effects are, at least in part, bacteria-selective. <i>E. coli</i> HB101 and <i>E. cloacae</i> TW1 exhibited increased adhesion/invasion onto small intestine epithelial cells when exposed to <i>Giardia</i> EVs. EVs labelled with PKH67 revealed colocalization with <i>E. coli</i> HB101 and <i>E. cloacae</i> TW1 bacterial cells. Small RNA sequencing revealed a high abundance of ribosomal RNA (rRNA)- and transfer RNA (tRNA)-derived small RNAs, short-interfering RNAs (siRNAs) and micro-RNAs (miRNAs) within <i>Giardia</i> EVs. Proteomic analysis of EVs uncovered the presence of RNA chaperones and heat shock proteins that can facilitate the thermal stability of EVs and its sRNA cargo, as well as protein-modifying enzymes. In vitro, RNase heat-treatment assays showed that total RNAs in EVs, but not proteins, are responsible for modulating bacterial swimming motility and biofilm formation. <i>G. duodenalis</i> small RNAs of EVs, but not proteins, were responsible for the increased bacterial adhesion to intestinal epithelial cells induced upon exposure to <i>Giardia</i> EVs. Together, the findings indicate that <i>Giardia</i> EVs contain a heat-stable, RNase-sensitive cargo that can trigger the development of pathobiont characteristics in Enterobacteria, depicting a novel trans-kingdom cross-talk in the gut.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.109","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43818241","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicle-mediated protein delivery to the liver","authors":"Nazma F. Ilahibaks,&nbsp;Marieke T. Roefs,&nbsp;Maike A. D. Brans,&nbsp;Christian Snijders Blok,&nbsp;Saskia C. A. de Jager,&nbsp;Raymond M. Schiffelers,&nbsp;Pieter Vader,&nbsp;Zhiyong Lei,&nbsp;Joost P. G. Sluijter","doi":"10.1002/jex2.97","DOIUrl":"10.1002/jex2.97","url":null,"abstract":"<p>Extracellular vesicles (EVs) are nanoscale particles that facilitate intercellular communication. They are regarded as a promising natural drug delivery system for transporting and delivering bioactive macromolecules to target cells. Recently, researchers have engineered EVs with FKBP12/FRB heterodimerization domains that interact with rapamycin to load and deliver exogenous proteins for both <i>in vitro</i> and <i>in vivo</i> applications. In this study, we examined the tissue distribution of EVs using near-infrared fluorescent imaging. We evaluated the effectiveness of EV-mediated delivery of Cre recombinase specifically to hepatocytes in the livers of Ai9 Cre-loxP reporter mice. Intravenous injection resulted in more efficient Cre protein delivery to the liver than intraperitoneal injections. Depleting liver-resident macrophages with clodronate-encapsulated liposome pre-treatment did not enhance EV-mediated Cre delivery to hepatocytes. Moreover, we demonstrated that multiple intravenous injections of Cre-EVs facilitated functional Cre delivery to hepatocytes. To the best of our knowledge, this is the first study to simultaneously investigate the tissue distribution of FKBP12/FRB-engineered EVs and their subsequent intracellular protein delivery in Ai9 Cre-loxP reporter mice. These insights can inform preclinical research and contribute to developing next-generation EV-based platforms for delivering therapeutic proteins or genome editing technologies targeting the liver.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 9","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-08-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.97","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44937477","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Survey of organ-derived small extracellular vesicles and particles (sEVPs) to identify selective protein markers in mouse serum","authors":"Kotb Abdelmohsen,&nbsp;Allison B. Herman,&nbsp;Angelica E. Carr,&nbsp;Charnae’ A. Henry-Smith,&nbsp;Martina Rossi,&nbsp;Qiong Meng,&nbsp;Jen-Hao Yang,&nbsp;Dimitrios Tsitsipatis,&nbsp;Alhassan Bangura,&nbsp;Rachel Munk,&nbsp;Jennifer L. Martindale,&nbsp;Carlos J. Nogueras-Ortiz,&nbsp;Jon Hao,&nbsp;Yi Gong,&nbsp;Yie Liu,&nbsp;Chang-Yi Cui,&nbsp;Lisa M. Hartnell,&nbsp;Nathan L. Price,&nbsp;Luigi Ferrucci,&nbsp;Dimitrios Kapogiannis,&nbsp;Rafael de Cabo,&nbsp;Myriam Gorospe","doi":"10.1002/jex2.106","DOIUrl":"10.1002/jex2.106","url":null,"abstract":"<p>Extracellular vesicles and particles (EVPs) are secreted by organs across the body into different circulatory systems, including the bloodstream, and reflect pathophysiologic conditions of the organ. However, the heterogeneity of EVPs in the blood makes it challenging to determine their organ of origin. We hypothesized that small (s)EVPs (&lt;100 nm in diameter) in the bloodstream carry distinctive protein signatures associated with each originating organ, and we investigated this possibility by studying the proteomes of sEVPs produced by six major organs (brain, liver, lung, heart, kidney, and fat). We found that each organ contained distinctive sEVP proteins: 68 proteins were preferentially found in brain sEVPs, 194 in liver, 39 in lung, 15 in heart, 29 in kidney, and 33 in fat. Furthermore, we isolated sEVPs from blood and validated the presence of sEVP proteins associated with the brain (DPP6, SYT1, DNM1L), liver (FABPL, ARG1, ASGR1/2), lung (SFPTA), heart (CPT1B), kidney (SLC31), and fat (GDN). We further discovered altered levels of these proteins in serum sEVPs obtained from old mice compared to young mice. In sum, we have cataloged sEVP proteins that can serve as potential biomarkers for organ identification in serum and show differential expression with age.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-08-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.106","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41164303","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A commentary on: TDO2-augmented fibroblasts secrete EVs enriched in immunomodulatory Y-derived small RNA","authors":"Clarissa Sastrawidjaya,&nbsp;Phuong H. D. Nguyen","doi":"10.1002/jex2.99","DOIUrl":"https://doi.org/10.1002/jex2.99","url":null,"abstract":"<p>Extracellular vesicles (EVs) containing biological information in the form of proteins, lipids, and nucleic acids can be shuttled between cells to mediate physiological responses (Raposo &amp; Stahl, <span>2019</span>). Functions of EVs in healthy states include intercellular communication, cell survival, angiogenesis, inflammatory and immune response, coagulation, and waste management (Yuana et al., <span>2013</span>). By the same token, EVs are involved in pathophysiological conditions. Alterations in EV cargo following changes to the pathophysiological state of donor cells correlate with disease progression (Saheera et al., <span>2021</span>). EVs have attracted immense interest in the field of cell-free therapeutics. Compared to cells, EVs are more stable, less immunogenic, and can tolerate repeated freeze–thaw cycles, making them less burdensome for manufacture and storage (Akers et al., <span>2016</span>; Murphy et al., <span>2019</span>; Usman et al., <span>2018</span>). Due to their nanoscale dimensions, EVs are in some instancescapable of permeating into tissues and crossing cellular barriers (Banks et al., <span>2020</span>; Saint-Pol et al., <span>2020</span>). They may therefore be more efficient in reaching target sites of action.</p><p>In the context of tissue injury, the ability of EVs to modulate the immune response suggests their therapeutic potential. Various studies have demonstrated the importance of EVs in restoring tissue homeostasis following acute injuries, trauma, surgeries, and chronic diseases (Arslan et al., <span>2013</span>; Gallet et al., <span>2016</span>; Kervadec et al., <span>2016</span>; Xiong et al., <span>2017</span>,<span>2020</span>). After all, immune responses crucially determine the prognosis of tissue injury, whereby outcomes range from fibrosis to complete regeneration depending on the type, duration, and cellular mediators underlying the immune response (Julier et al., <span>2017</span>). The present commentary addresses the findings of Ciullo et al. that a small Y-derived RNA, NT4, mediates immunomodulatory effects when enriched in EVs secreted by tryptophan 2,3-dioxygenase (TDO2)-upregulated fibroblasts (TDO2-EVs) (Ciullo et al., <span>2023</span>) (Figure 1). NT4 enrichment allows TDO2-EVs to attenuate inflammatory profiles in macrophages, contributing to cardioprotection in disease states such as acute myocardial infarction (AMI).</p><p>In myocardial infarction (MI), myocardial tissues undergo ischemic death, resulting in a large-scale loss of cardiac muscle including cardiomyocytes, endothelial cells, and fibroblasts (Laflamme &amp; Murry, <span>2005</span>). Depending on the severity of the damage, patients may suffer from progressive contractile dysfunction. Following MI, a process of repair, known as ventricular remodelling, stimulates a vigorous inflammatory response to recruit immune cells to the injured site for clearing apoptotic cells and debris. However, unrestrained inflammation post","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-08-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.99","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50151963","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TP53 mutations correlate with the non-coding RNA content of small extracellular vesicles in melanoma","authors":"Maureen Labbé,&nbsp;Estelle Menoret,&nbsp;Franck Letourneur,&nbsp;Benjamin Saint-Pierre,&nbsp;Laurence de Beaurepaire,&nbsp;Joëlle Veziers,&nbsp;Brigitte Dreno,&nbsp;Marc G. Denis,&nbsp;Christophe Blanquart,&nbsp;Nicolas Boisgerault,&nbsp;Jean-François Fonteneau,&nbsp;Delphine Fradin","doi":"10.1002/jex2.105","DOIUrl":"10.1002/jex2.105","url":null,"abstract":"<p>Non-coding RNAs (ncRNAs) are important regulators of gene expression. They are expressed not only in cells, but also in cell-derived extracellular vesicles (EVs). The mechanisms controlling their loading and sorting remain poorly understood. Here, we investigated the impact of <i>TP53</i> mutations on the non-coding RNA content of small melanoma EVs. After purification of small EVs from six different patient-derived melanoma cell lines, we characterized them by small RNA sequencing and lncRNA microarray analysis. We found that <i>TP53</i> mutations are associated with a specific micro and long non-coding RNA content in small EVs. Then, we showed that long and small non-coding RNAs enriched in <i>TP53</i> mutant small EVs share a common sequence motif, highly similar to the RNA-binding motif of Sam68, a protein interacting with hnRNP proteins. This protein thus may be an interesting partner of p53, involved in the expression and loading of the ncRNAs. To conclude, our data support the existence of cellular mechanisms associate with <i>TP53</i> mutations which control the ncRNA content of small EVs in melanoma.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.105","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41510826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Adipose derived stem cell extracellular vesicles modulate primary human macrophages to an anti-inflammatory phenotype in vitro","authors":"Emma K. C. Symonds,&nbsp;Bianca Black,&nbsp;Alexander Brown,&nbsp;Ineke Meredith,&nbsp;Margaret J. Currie,&nbsp;Kathryn E. Hally,&nbsp;Kirsty M. Danielson","doi":"10.1002/jex2.104","DOIUrl":"10.1002/jex2.104","url":null,"abstract":"<p>EVs released by adipose derived stem cells (ADSCs) have shown promise as a therapeutic for tissue repair because of their purported immune-regulatory properties. Extracellular vesicles (EVs) from ADSCs could be beneficial in improving graft retention rates for autologous fat grafting (AFG) post-mastectomy as, currently, grafted tissue rates are variable. Enriching grafted tissue with ADSC-EVs may improve retention rates by modulating macrophages resident within both the breast and lipoaspirate. We aimed to identify key macrophage phenotypes that are modulated by ADSC-EVs in vitro. ADSCs were isolated from lipoaspirates of women undergoing AFG and characterised by flow cytometry and differentiation potential. ADSC-EVs were isolated from culture media and characterised by tuneable resistive pulse sensing, transmission electron microscopy and Western blot. Primary monocyte-derived macrophages were polarized to an M1-like (GM-CSF, IFNγ), M2-like phenotype (M-CSF, IL-4) or maintained (M0-like; M-CSF) and ADSC-EVs were co-cultured with macrophages for 48 h. Flow cytometry and high-dimensional analysis clustered macrophages post co-culture. A manual gating strategy was generated to recapitulate these clusters and was applied to a repeat experimental run. Both runs were analysed to examine the prevalence of each cluster, representing a unique macrophage phenotype, with and without ADSC-EVs. Following the addition of ADSC-EVs, M0-like macrophages demonstrated a reciprocal shift of cell distribution from a cluster with a ‘high inflammatory profile’ (CD36⁺⁺⁺CD206⁺⁺⁺CD86⁺⁺⁺; 16.5 ± 7.0%; <i>p</i> &lt; 0.0001) to a cluster with a ‘lower inflammatory profile’ (CD36^+CD206+CD86+; 35  ± 21.5%; <i>p</i> &lt; 0.05). M1-like macrophages shifted from a cluster with a ‘high inflammatory profile’ (CD206⁺⁺CD11b⁺⁺CD36⁺⁺CD163⁺⁺; 26.1 ± 9.4%; <i>p</i> = 0.0024) to a ‘lower inflammatory profile’ (CD206⁺CD11b⁺CD36^+CD163+; 72.8  ± 8.7%; <i>p</i> = 0.0007). There was no shift in M2-like clusters following ADSC-EV treatment. ADSC-EVs are complex regulators of macrophage phenotype that can shift macrophages away from a heightened pro-inflammatory state.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 8","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.104","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"49239880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Internalization of extracellular vesicles from Lactobacillus johnsonii N6.2 elicit an RNA sensory response in human pancreatic cell lines","authors":"Danilo R. da Silva,&nbsp;Claudio F. Gonzalez,&nbsp;Graciela Lorca","doi":"10.1002/jex2.101","DOIUrl":"https://doi.org/10.1002/jex2.101","url":null,"abstract":"<p>Cells of all domains of life can secrete extracellular vesicles (EV). These secreted vesicles have been indicated as vehicles carrying molecules that facilitate intra- and inter-species interaction. <i>Lactobacillus johnsonii</i> N6.2, a bacterium used in probiotic preparations, has been shown to produce nano-sized EV. In the present work we used <i>L. johnsonii</i> N6.2 EV, concentrated from exosome-depleted MRS supernatant, to identify the uptake mechanisms of EV and the impact of the RNA cargo in the EV on the upregulation of the cellular response of βlox5 human pancreatic cells. Using eukaryotic uptake inhibitors, it was found that EV are internalized by the clathrin/dynamin mediated endocytosis pathway. Further co-localization experiments with the endosome markers RAB5, RAB7 and LAMP1 as well as calcein indicated that EV escape the endosome shortly after RAB7 fusion. Using the expression of the 2′,5′-oligoadenylate synthetase (OAS) host pathway, previously identified as targeted by <i>L. johnsonii</i> EV, we found that the host cellular response to the EV are dependent on the integrity of the external components of the EV as well as on the RNA cargo. Global transcriptome analysis was performed on EV and the bacterial whole cell. It was found that the RNA transcripts found within the EV largely represent the most abundantly transcribed genes in the bacterial cells such as those associated with protein synthesis and glycolysis. Further analysis showed an enrichment of smaller size transcripts as well as those encoding for membrane bound or extracellular proteins in <i>L. johnsonii</i>’s EV.</p>","PeriodicalId":73747,"journal":{"name":"Journal of extracellular biology","volume":"2 7","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-07-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jex2.101","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"50136936","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}