Journal of Extracellular Vesicles最新文献

{"title":"Extracellular vesicles containing SARS-CoV-2 proteins are associated with multi-organ dysfunction and worse outcomes in patients with severe COVID-19","authors":"Diego de Miguel-Perez,&nbsp;Marisol Arroyo-Hernandez,&nbsp;Sabrina La Salvia,&nbsp;Muthukumar Gunasekaran,&nbsp;Edward M. Pickering,&nbsp;Stephanie Avila,&nbsp;Etse Gebru,&nbsp;Eduardo Becerril-Vargas,&nbsp;Sergio Monraz-Perez,&nbsp;Kapil Saharia,&nbsp;Alison Grazioli,&nbsp;Michael T. McCurdy,&nbsp;Matthew Frieman,&nbsp;Lisa Miorin,&nbsp;Alessandro Russo,&nbsp;Andrés F. Cardona,&nbsp;Adolfo García-Sastre,&nbsp;Sunjay Kaushal,&nbsp;Fred R. Hirsch,&nbsp;Djordje Atanackovic,&nbsp;Susmita Sahoo,&nbsp;Oscar Arrieta,&nbsp;Christian Rolfo","doi":"10.1002/jev2.70001","DOIUrl":"https://doi.org/10.1002/jev2.70001","url":null,"abstract":"<p>The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19) and has been related to more than 7 million deaths globally since 2019. The association of high levels of IL-6 with severe cases led to the early evaluation of the anti-IL6 inhibitor tocilizumab as a potential treatment, which unfortunately failed to improve survival in many trials. Moreover, little is known about the development of COVID-19 sequelae, and biomarkers are needed to understand and anticipate these processes. Because extracellular vesicles (EVs) play an important role in viral infection and immune response, they could potentially serve as predictive and prognostic biomarkers. We isolated EVs from 39 patients with severe COVID-19, from which 29 received tocilizumab and 10 were considered controls. Blood samples, which were collected at hospitalisation before treatment, at Day 7, and Day 15 during follow-up, were assessed by immunoblot for longitudinal expression of spike (S) and nucleocapsid (N) proteins. Dynamic expression was calculated and compared with clinicopathological and experimental variables. Expression of EV S was validated by immunogold and imaging flow-cytometry, revealing an enrichment in CD9+ EVs. As a result, decreasing expression of EV viral proteins was observed in patients treated with tocilizumab. Moreover, higher increase in EV S was observed in patients with lower antibody response, hyperfibrinogenemia, lower respiratory function, higher blood pressure and shorter outcomes. These findings lay the foundation for future studies characterizing the role of EVs in multiorgan assessment and identifying biomarkers in patients with severe COVID-19 and possible long COVID.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 11","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-11-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.70001","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142666152","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":"Efficient enzyme-free isolation of brain-derived extracellular vesicles","authors":"Andreu Matamoros-Angles,&nbsp;Emina Karadjuzovic,&nbsp;Behnam Mohammadi,&nbsp;Feizhi Song,&nbsp;Santra Brenna,&nbsp;Susanne Caroline Meister,&nbsp;Bente Siebels,&nbsp;Hannah Voß,&nbsp;Carolin Seuring,&nbsp;Isidre Ferrer,&nbsp;Hartmut Schlüter,&nbsp;Matthias Kneussel,&nbsp;Hermann Clemens Altmeppen,&nbsp;Michaela Schweizer,&nbsp;Berta Puig,&nbsp;Mohsin Shafiq,&nbsp;Markus Glatzel","doi":"10.1002/jev2.70011","DOIUrl":"10.1002/jev2.70011","url":null,"abstract":"<p>Extracellular vesicles (EVs) have gained significant attention as pathology mediators and potential diagnostic tools for neurodegenerative diseases. However, isolation of brain-derived EVs (BDEVs) from tissue remains challenging, often involving enzymatic digestion steps that may compromise the integrity of EV proteins and overall functionality. Here, we describe that collagenase digestion, commonly used for BDEV isolation, produces undesired protein cleavage of EV-associated proteins in brain tissue homogenates and cell-derived EVs. In order to avoid this effect, we studied the possibility of isolating BDEVs with a reduced amount of collagenase or without any protease. Characterization of the isolated BDEVs from mouse and human samples (both female and male) revealed their characteristic morphology and size distribution with both approaches. However, we show that even minor enzymatic digestion induces ‘artificial’ proteolytic processing in key BDEV markers, such as Flotillin-1, CD81, and the cellular prion protein (PrP^C), whereas avoiding enzymatic treatment completely preserves their integrity. We found no major differences in mRNA and protein content between non-enzymatically and enzymatically isolated BDEVs, suggesting that the same BDEV populations are purified with both approaches. Intriguingly, the lack of Golgi marker GM130 signal, often referred to as contamination indicator (or negative marker) in EV preparations, seems to result from enzymatic digestion rather than from its actual absence in BDEV samples. Overall, we show that non-enzymatic isolation of EVs from brain tissue is possible and avoids artificial pruning of proteins while achieving an overall high BDEV yield and purity. This protocol will help to understand the functions of BDEV and their associated proteins in a near-physiological setting, thus opening new research approaches.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 11","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11541858/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604686","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":"Hypoxia and TNF-alpha modulate extracellular vesicle release from human induced pluripotent stem cell-derived cardiomyocytes","authors":"Margarida Viola,&nbsp;Maarten P. Bebelman,&nbsp;Renee G. C. Maas,&nbsp;Willemijn S. de Voogt,&nbsp;Frederik J. Verweij,&nbsp;Cor S. Seinen,&nbsp;Saskia C. A. de Jager,&nbsp;Pieter Vader,&nbsp;Dirk Michiel Pegtel,&nbsp;Joost Petrus Gerardus Sluijter","doi":"10.1002/jev2.70000","DOIUrl":"10.1002/jev2.70000","url":null,"abstract":"<p>Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in the heart under homeostatic and pathological conditions, such as myocardial infarction (MI). However, the basic mechanisms driving cardiomyocyte-derived EV (CM-EV) production following stress are poorly understood. In this study, we generated human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) that express NanoLuc-tetraspanin reporters. These modified hiPSC-CMs allow for quantification of tetraspanin-positive CM-EV secretion from small numbers of cells without the need for time-consuming EV isolation techniques. We subjected these cells to a panel of small molecules to study their effect on CM-EV biogenesis and secretion under basal and stress-associated conditions. We observed that EV biogenesis is context-dependent in hiPSC-CMs. Nutrient starvation decreases CM-EV secretion while hypoxia increases the production of CM-EVs in a nSmase2-dependent manner. Moreover, the inflammatory cytokine TNF-α increased CM-EV secretion through a process involving NLRP3 inflammasome activation and mTOR signalling. Here, we detailed for the first time the regulatory mechanisms of EV biogenesis in hiPSC-CMs upon MI-associated stressors.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 11","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11541862/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604691","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":"PlexinA1 (PLXNA1) as a novel scaffold protein for the engineering of extracellular vesicles","authors":"Hang Zhao,&nbsp;Zhi Li,&nbsp;Da Liu,&nbsp;Jiaxun Zhang,&nbsp;Zhicheng You,&nbsp;Yuzhang Shao,&nbsp;Hongyan Li,&nbsp;Jun Yang,&nbsp;Xiang Liu,&nbsp;Miaozhu Wang,&nbsp;Chengen Wu,&nbsp;Jing Chen,&nbsp;Jianwu Wang,&nbsp;Guanyi Kong,&nbsp;Libo Zhao","doi":"10.1002/jev2.70012","DOIUrl":"10.1002/jev2.70012","url":null,"abstract":"<p>Extracellular vesicles (EVs) had been described as a next-generation drug delivery system, due to the compelling evidence that they can facilitate the transfer of a variety of biomolecules between cells. The most frequently used strategy for loading protein cargoes is the endogenous engineering of EVs through genetic fusion of the protein of interest (POI) and scaffold proteins with high EV-sorting ability. However, the lack of scaffold proteins had become a major issue hindering the promotion of this technology. Herein, we proposed novel screening criteria that relax the inclusion requirement of candidate scaffold proteins and eventually identified a new scaffold protein, PLXNA1. The truncated PLXNA1 not only inherits the high EV-sorting ability of its full-length counterpart but also allows the fusion expression of POI in both outer surface and luminal areas, individually or simultaneously. In conclusion, our screening criteria expanded the range of potential scaffold proteins. The identified scaffold protein PLXNA1 showed great potential in developing therapeutic EVs.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 11","pages":""},"PeriodicalIF":15.5,"publicationDate":"2024-11-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11541859/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142604712","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 containing SARS-CoV-2 proteins are associated with multi-organ dysfunction and worse outcomes in patients with severe COVID-19.","authors":"Diego de Miguel-Perez, Marisol Arroyo-Hernandez, Sabrina La Salvia, Muthukumar Gunasekaran, Edward M Pickering, Stephanie Avila, Etse Gebru, Eduardo Becerril-Vargas, Sergio Monraz-Perez, Kapil Saharia, Alison Grazioli, Michael T McCurdy, Matthew Frieman, Lisa Miorin, Alessandro Russo, Andrés F Cardona, Adolfo García-Sastre, Sunjay Kaushal, Fred R Hirsch, Djordje Atanackovic, Susmita Sahoo, Oscar Arrieta, Christian Rolfo","doi":"10.1002/jev2.70001","DOIUrl":"https://doi.org/10.1002/jev2.70001","url":null,"abstract":"<p><p>The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) causes coronavirus disease 2019 (COVID-19) and has been related to more than 7 million deaths globally since 2019. The association of high levels of IL-6 with severe cases led to the early evaluation of the anti-IL6 inhibitor tocilizumab as a potential treatment, which unfortunately failed to improve survival in many trials. Moreover, little is known about the development of COVID-19 sequelae, and biomarkers are needed to understand and anticipate these processes. Because extracellular vesicles (EVs) play an important role in viral infection and immune response, they could potentially serve as predictive and prognostic biomarkers. We isolated EVs from 39 patients with severe COVID-19, from which 29 received tocilizumab and 10 were considered controls. Blood samples, which were collected at hospitalisation before treatment, at Day 7, and Day 15 during follow-up, were assessed by immunoblot for longitudinal expression of spike (S) and nucleocapsid (N) proteins. Dynamic expression was calculated and compared with clinicopathological and experimental variables. Expression of EV S was validated by immunogold and imaging flow-cytometry, revealing an enrichment in CD9+ EVs. As a result, decreasing expression of EV viral proteins was observed in patients treated with tocilizumab. Moreover, higher increase in EV S was observed in patients with lower antibody response, hyperfibrinogenemia, lower respiratory function, higher blood pressure and shorter outcomes. These findings lay the foundation for future studies characterizing the role of EVs in multiorgan assessment and identifying biomarkers in patients with severe COVID-19 and possible long COVID.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"13 11","pages":"e70001"},"PeriodicalIF":15.5,"publicationDate":"2024-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142668154","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":1,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Physical association of low density lipoprotein particles and extracellular vesicles unveiled by single particle analysis","authors":"Estefanía Lozano-Andrés,&nbsp;Agustin Enciso-Martinez,&nbsp;Abril Gijsbers,&nbsp;Andrea Ridolfi,&nbsp;Guillaume Van Niel,&nbsp;Sten F. W. M. Libregts,&nbsp;Cláudio Pinheiro,&nbsp;Martijn J. C. van Herwijnen,&nbsp;An Hendrix,&nbsp;Marco Brucale,&nbsp;Francesco Valle,&nbsp;Peter J. Peters,&nbsp;Cees Otto,&nbsp;Ger J. A. Arkesteijn,&nbsp;Marca H. M. Wauben","doi":"10.1002/jev2.12376","DOIUrl":"10.1002/jev2.12376","url":null,"abstract":"<p>Extracellular vesicles (EVs) in blood plasma are recognized as potential biomarkers for disease. Although blood plasma is easily obtainable, analysis of EVs at the single particle level is still challenging due to the biological complexity of this body fluid. Besides EVs, plasma contains different types of lipoproteins particles (LPPs), that outnumber EVs by orders of magnitude and which partially overlap in biophysical properties such as size, density and molecular makeup. Consequently, during EV isolation LPPs are often co-isolated. Furthermore, physical EV-LPP complexes have been observed in purified EV preparations. Since co-isolation or association of LPPs can impact EV-based analysis and biomarker profiling, we investigated the presence and formation of EV-LPP complexes in biological samples by using label-free atomic force microscopy, cryo-electron tomography and synchronous Rayleigh and Raman scattering analysis of optically trapped particles and fluorescence-based high sensitivity single particle flow cytometry. Furthermore, we evaluated the impact on flow cytometric analysis in the presence of LPPs using in vitro spike-in experiments of purified tumour cell line-derived EVs in different classes of purified human LPPs. Based on orthogonal single-particle analysis techniques we demonstrate that EV-LPP complexes can form under physiological conditions. Furthermore, we show that in fluorescence-based flow cytometric EV analysis staining of LPPs, as well as EV-LPP associations, can influence quantitative and qualitative EV analysis. Lastly, we demonstrate that the colloidal matrix of the biofluid in which EVs reside impacts their buoyant density, size and/or refractive index (RI), which may have consequences for down-stream EV analysis and EV biomarker profiling.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"12 11","pages":""},"PeriodicalIF":16.0,"publicationDate":"2023-11-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10634195/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71521726","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":"RTP801 mediates transneuronal toxicity in culture via extracellular vesicles","authors":"Júlia Solana-Balaguer,&nbsp;Núria Martín-Flores,&nbsp;Pol Garcia-Segura,&nbsp;Genís Campoy-Campos,&nbsp;Leticia Pérez-Sisqués,&nbsp;Almudena Chicote-González,&nbsp;Joaquín Fernández-Irigoyen,&nbsp;Enrique Santamaría,&nbsp;Esther Pérez-Navarro,&nbsp;Jordi Alberch,&nbsp;Cristina Malagelada","doi":"10.1002/jev2.12378","DOIUrl":"10.1002/jev2.12378","url":null,"abstract":"<p>Extracellular vesicles (EVs) play a crucial role in intercellular communication, participating in the paracrine trophic support or in the propagation of toxic molecules, including proteins. RTP801 is a stress-regulated protein, whose levels are elevated during neurodegeneration and induce neuron death. However, whether RTP801 toxicity is transferred trans-neuronally via EVs remains unknown. Hence, we overexpressed or silenced RTP801 protein in cultured cortical neurons, isolated their derived EVs (RTP801-EVs or shRTP801-EVs, respectively), and characterized EVs protein content by mass spectrometry (MS). RTP801-EVs toxicity was assessed by treating cultured neurons with these EVs and quantifying apoptotic neuron death and branching. We also tested shRTP801-EVs functionality in the pathologic in vitro model of 6-Hydroxydopamine (6-OHDA). Expression of RTP801 increased the number of EVs released by neurons. Moreover, RTP801 led to a distinct proteomic signature of neuron-derived EVs, containing more pro-apoptotic markers. Hence, we observed that RTP801-induced toxicity was transferred to neurons via EVs, activating apoptosis and impairing neuron morphology complexity. In contrast, shRTP801-EVs were able to increase the arborization in recipient neurons. The 6-OHDA neurotoxin elevated levels of RTP801 in EVs, and 6-OHDA-derived EVs lost the mTOR/Akt signalling activation via Akt and RPS6 downstream effectors. Interestingly, EVs derived from neurons where RTP801 was silenced prior to exposing them to 6-OHDA maintained Akt and RPS6 transactivation in recipient neurons. Taken together, these results suggest that RTP801-induced toxicity is transferred via EVs, and therefore, it could contribute to the progression of neurodegenerative diseases, in which RTP801 is involved.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"12 11","pages":""},"PeriodicalIF":16.0,"publicationDate":"2023-11-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10627824/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71482314","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":"Engineering a tunable micropattern-array assay to sort single extracellular vesicles and particles to detect RNA and protein in situ","authors":"Jingjing Zhang,&nbsp;Xilal Y. Rima,&nbsp;Xinyu Wang,&nbsp;Luong T. H. Nguyen,&nbsp;Kristin Huntoon,&nbsp;Yifan Ma,&nbsp;Paola Loreto Palacio,&nbsp;Kim Truc Nguyen,&nbsp;Karunya Albert,&nbsp;Minh-Dao Duong-Thi,&nbsp;Nicole Walters,&nbsp;Kwang Joo Kwak,&nbsp;Min Jin Yoon,&nbsp;Hong Li,&nbsp;Jacob Doon-Ralls,&nbsp;Colin L. Hisey,&nbsp;Daeyong Lee,&nbsp;Yifan Wang,&nbsp;Jonghoon Ha,&nbsp;Kelsey Scherler,&nbsp;Shannon Fallen,&nbsp;Inyoul Lee,&nbsp;Andre F. Palmer,&nbsp;Wen Jiang,&nbsp;Setty M. Magaña,&nbsp;Kai Wang,&nbsp;Betty Y. S. Kim,&nbsp;L. James Lee,&nbsp;Eduardo Reátegui","doi":"10.1002/jev2.12369","DOIUrl":"10.1002/jev2.12369","url":null,"abstract":"<p>The molecular heterogeneity of extracellular vesicles (EVs) and the co-isolation of physically similar particles, such as lipoproteins (LPs), confounds and limits the sensitivity of EV bulk biomarker characterization. Herein, we present a single-EV and particle (siEVP) protein and RNA assay (^siEVPPRA) to simultaneously detect mRNAs, miRNAs, and proteins in subpopulations of EVs and LPs. The ^siEVPPRA immobilizes and sorts particles via positive immunoselection onto micropatterns and focuses biomolecular signals in situ. By detecting EVPs at a single-particle resolution, the ^siEVPPRA outperformed the sensitivities of bulk-analysis benchmark assays for RNA and protein. To assess the specificity of RNA detection in complex biofluids, EVs from various glioma cell lines were processed with small RNA sequencing, whereby two mRNAs and two miRNAs associated with glioblastoma multiforme (GBM) were chosen for cross-validation. Despite the presence of single-EV-LP co-isolates in serum, the ^siEVPPRA detected GBM-associated vesicular RNA profiles in GBM patient siEVPs. The ^siEVPPRA effectively examines intravesicular, intervesicular, and interparticle heterogeneity with diagnostic promise.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"12 11","pages":""},"PeriodicalIF":16.0,"publicationDate":"2023-10-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10618633/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71424073","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":"Palmitate and glucose increase amyloid precursor protein in extracellular vesicles: Missing link between metabolic syndrome and Alzheimer's disease","authors":"Bhumsoo Kim,&nbsp;Yoon-Tae Kang,&nbsp;Faye E. Mendelson,&nbsp;John M. Hayes,&nbsp;Masha G. Savelieff,&nbsp;Sunitha Nagrath,&nbsp;Eva L. Feldman","doi":"10.1002/jev2.12340","DOIUrl":"10.1002/jev2.12340","url":null,"abstract":"<p>The metabolic syndrome (MetS) and Alzheimer's disease share several pathological features, including insulin resistance, abnormal protein processing, mitochondrial dysfunction and elevated inflammation and oxidative stress. The MetS constitutes elevated fasting glucose, obesity, dyslipidaemia and hypertension and increases the risk of developing Alzheimer's disease, but the precise mechanism remains elusive. Insulin resistance, which develops from a diet rich in sugars and saturated fatty acids, such as palmitate, is shared by the MetS and Alzheimer's disease. Extracellular vesicles (EVs) are also a point of convergence, with altered dynamics in both the MetS and Alzheimer's disease. However, the role of palmitate- and glucose-induced insulin resistance in the brain and its potential link through EVs to Alzheimer's disease is unknown. We demonstrate that palmitate and high glucose induce insulin resistance and amyloid precursor protein phosphorylation in primary rat embryonic cortical neurons and human cortical stem cells. Palmitate also triggers insulin resistance in oligodendrocytes, the supportive glia of the brain. Palmitate and glucose enhance amyloid precursor protein secretion from cortical neurons via EVs, which induce tau phosphorylation when added to naïve neurons. Additionally, EVs from palmitate-treated oligodendrocytes enhance insulin resistance in recipient neurons. Overall, our findings suggest a novel theory underlying the increased risk of Alzheimer's disease in MetS mediated by EVs, which spread Alzheimer's pathology and insulin resistance.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"12 11","pages":""},"PeriodicalIF":16.0,"publicationDate":"2023-10-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10613125/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"66783937","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":"Culture conditions greatly impact the levels of vesicular and extravesicular Ago2 and RNA in extracellular vesicle preparations","authors":"Lizandra Jimenez,&nbsp;Bahnisikha Barman,&nbsp;Youn Jae Jung,&nbsp;Lauren Cocozza,&nbsp;Evan Krystofiak,&nbsp;Cherie Saffold,&nbsp;Kasey C. Vickers,&nbsp;John T. Wilson,&nbsp;T. Renee Dawson,&nbsp;Alissa M. Weaver","doi":"10.1002/jev2.12366","DOIUrl":"10.1002/jev2.12366","url":null,"abstract":"<p>Extracellular vesicle (EV)-carried miRNAs can influence gene expression and functional phenotypes in recipient cells. Argonaute 2 (Ago2) is a key miRNA-binding protein that has been identified in EVs and could influence RNA silencing. However, Ago2 is in a non-vesicular form in serum and can be an EV contaminant. In addition, RNA-binding proteins (RBPs), including Ago2, and RNAs are often minor EV components whose sorting into EVs may be regulated by cell signaling state. To determine the conditions that influence detection of RBPs and RNAs in EVs, we evaluated the effect of growth factors, oncogene signaling, serum, and cell density on the vesicular and nonvesicular content of Ago2, other RBPs, and RNA in small EV (SEV) preparations. Media components affected both the intravesicular and extravesicular levels of RBPs and miRNAs in EVs, with serum contributing strongly to extravesicular miRNA contamination. Furthermore, isolation of EVs from hollow fiber bioreactors revealed complex preparations, with multiple EV-containing peaks and a large amount of extravesicular Ago2/RBPs. Finally, KRAS mutation impacts the detection of intra- and extra-vesicular Ago2. These data indicate that multiple cell culture conditions and cell states impact the presence of RBPs in EV preparations, some of which can be attributed to serum contamination.</p>","PeriodicalId":15811,"journal":{"name":"Journal of Extracellular Vesicles","volume":"12 11","pages":""},"PeriodicalIF":16.0,"publicationDate":"2023-10-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/jev2.12366","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"54229366","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}