Extracellular vesicles and circulating nucleic acids最新文献_第5页

Advances in extracellular vesicle isolation methods: a path towards cell-type specific EV isolation 细胞外囊泡分离方法的进展:通往细胞类型特异性EV分离的途径

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2023.14

Adnan Shami-Shah, Benjamin G Travis, David R. Walt

{"title":"Advances in extracellular vesicle isolation methods: a path towards cell-type specific EV isolation","authors":"Adnan Shami-Shah, Benjamin G Travis, David R. Walt","doi":"10.20517/evcna.2023.14","DOIUrl":"https://doi.org/10.20517/evcna.2023.14","url":null,"abstract":"Extracellular vesicles are small, heterogenous, phospholipid-rich vesicles that are secreted by all cells into the extracellular space. They play a key role in intercellular communication because they can transport a variety of biomolecules such as proteins, lipids, and nucleic acids between cells. As categorized by the International Society of Extracellular Vesicles (ISEV), the term EV encompasses different sub-types, including exosomes, microvesicles, and apoptotic bodies, which differ in their size, origin, and cargo. EVs can be isolated from biological fluids such as blood, urine, and cerebrospinal fluid, and their biomolecular content can be analyzed to monitor the progression of certain diseases. Therefore, EVs can be used as a new source of liquid biomarkers for advancing novel diagnostic and therapeutic tools. Isolating and analyzing EVs can be challenging due to their nanoscopic size and low abundance. Several techniques have been developed for the isolation and characterization of EVs, including ultracentrifugation, density gradient separation, size-exclusion chromatography, microfluidics, and magnetic bead-based/affinity methods. This review highlights advances in EV isolation techniques in the last decade and provides a perspective on their advantages, limitations, and potential application to cell-type specific EV isolation in the future.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"83806732","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Non-invasive prenatal diagnosis (NIPD): current and emerging technologies 无创产前诊断(NIPD):当前和新兴技术

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2022.44

B. Hanson, B. Paternoster, Nikita Povarnitsyn, E. Scotchman, L. Chitty, N. Chandler

{"title":"Non-invasive prenatal diagnosis (NIPD): current and emerging technologies","authors":"B. Hanson, B. Paternoster, Nikita Povarnitsyn, E. Scotchman, L. Chitty, N. Chandler","doi":"10.20517/evcna.2022.44","DOIUrl":"https://doi.org/10.20517/evcna.2022.44","url":null,"abstract":"Prenatal testing is important for the early detection and diagnosis of rare genetic conditions with life-changing implications for the patient and their family. Gaining access to the fetal genotype can be achieved using gold-standard invasive sampling methods, such as amniocentesis and chorionic villus sampling, but these carry a small risk of miscarriage. Non-invasive prenatal diagnosis (NIPD) for select rare monogenic conditions has been in clinical service in England since 2012 and has revolutionised the field of prenatal diagnostics by reducing the number of women undergoing invasive sampling procedures. Fetal-derived genomic material is present in a highly fragmented form amongst the maternal cell-free DNA (cfDNA) in circulation, with sequence coverage across the entire fetal genome. Cell-free fetal DNA (cffDNA) is the foundation for NIPD, and several technologies have been clinically implemented for the detection of paternally inherited and de novo pathogenic variants. Conversely, a low abundance of cffDNA within a high background of maternal cfDNA makes assigning maternally inherited variants to the fetal fraction a significantly more challenging task. Research is ongoing to expand available tests for maternal inheritance to include a broader range of monogenic conditions, as well as to uncover novel diagnostic avenues. This review covers the scope of technologies currently clinically available for NIPD of monogenic conditions and those still in the research pipeline towards implementation in the future.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"87386427","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Emerging evidence for dysregulated proteome cargoes of tau-propagating extracellular vesicles driven by familial mutations of tau and presenilin. 新证据表明，在家族性 tau 和 presenilin 基因突变的驱动下，tau-propagating 细胞外囊泡的蛋白质组载货量失调。

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 Epub Date: 2023-11-21 DOI: 10.20517/evcna.2023.44

Vivian Hook, Sonia Podvin, Charles Mosier, Ben Boyarko, Laura Seyffert, Haley Stringer, Robert A Rissman

{"title":"Emerging evidence for dysregulated proteome cargoes of tau-propagating extracellular vesicles driven by familial mutations of tau and presenilin.","authors":"Vivian Hook, Sonia Podvin, Charles Mosier, Ben Boyarko, Laura Seyffert, Haley Stringer, Robert A Rissman","doi":"10.20517/evcna.2023.44","DOIUrl":"10.20517/evcna.2023.44","url":null,"abstract":"Tau propagation, pathogenesis, and neurotoxicity are hallmarks of neurodegenerative diseases that result in cognitive impairment. Tau accumulates in Alzheimer's disease (AD), frontotemporal dementia and parkinsonism linked to chromosome 17 (FTDP-17), chronic traumatic encephalopathy (CTE), progressive supranuclear palsy, and related tauopathies. Knowledge of the mechanisms for tau propagation in neurodegeneration is necessary for understanding the development of dementia. Exosomes, known as extracellular vesicles (EVs), have emerged as participants in promoting tau propagation. Recent findings show that EVs generated by neurons expressing familial mutations of tauopathies of FTDP-17 (P301L and V337M) (mTau) and presenilin (A246E) (mPS1) in AD induce tau propagation and accumulation after injection into rodent brain. To gain knowledge of the proteome cargoes of the mTau and mPS1 EVs that promote tau pathogenesis, this review compares the proteomes of these EVs, which results in important new questions concerning EV mechanisms of tau pathogenesis. Proteomics data show that EVs produced by mTau- and mPS1-expressing iPSC neurons share proteins involved in exocytosis and vesicle secretion and, notably, these EVs also possess differences in protein components of vesicle-mediated transport, extracellular functions, and cell adhesion. It will be important for future studies to gain an understanding of the breadth of familial genetic mutations of tau, presenilin, and other genes in promoting EV initiation of tau propagation and pathogenesis. Furthermore, elucidation of EV cargo components that mediate tau propagation will have potential as biomarkers and therapeutic strategies to ameliorate dementia of tauopathies.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10732590/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138833346","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

Immunomodulatory role of EV-derived non-coding RNA in lung cancer ev衍生的非编码RNA在肺癌中的免疫调节作用

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2022.42

Patrizia Ghidotti, Ilaria Petraroia, O. Fortunato, F. Pontis

引用次数: 0

Detection by super-resolution microscopy of viral proteins inside bloodborne extracellular vesicles 血源性细胞外囊泡内病毒蛋白的超分辨显微镜检测

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2023.46

Rakesh K. Singh, Mark F. Santos, Charles Herndon, Brandon A. Gieler, Isaac Lee, Jiahui Chen, Aurelio Lorico

{"title":"Detection by super-resolution microscopy of viral proteins inside bloodborne extracellular vesicles","authors":"Rakesh K. Singh, Mark F. Santos, Charles Herndon, Brandon A. Gieler, Isaac Lee, Jiahui Chen, Aurelio Lorico","doi":"10.20517/evcna.2023.46","DOIUrl":"https://doi.org/10.20517/evcna.2023.46","url":null,"abstract":"Aim: Extracellular vesicles (EVs) are small particles released by all cells, including virally infected cells, into the extracellular space. They play a role in various cellular processes, including intercellular communication, signaling, and immunity, and carry several biomolecules like proteins, lipids, and nucleic acids that can modulate cellular functions mostly by releasing their cargo inside the target cells via the endocytic pathway. One of the most exciting aspects of EV physiology is its potential in liquid biopsy as a diagnostic and prognostic marker. However, due to their extremely small size and lack of a molecular approach to examine intravesicular content or cargo, we cannot fully utilize their potential in healthcare. Methods: Here, we present a novel approach that allows examining bloodborne EVs at a single-particle level with the ability to examine their cargo without disrupting structural integrity. Our technique utilizes super-resolution microscopy and a unique permeabilization process that maintains structural integrity while facilitating the examination of EV cargo. We used a mild-detergent-based permeabilization buffer that protects the integrity of EVs, minimizes background, and improves detection. Results: Utilizing this approach, we were able to recognize viral proteins of SARS-CoV-2 virus in COVID-19 patients, including spike and nucleocapsid. Surprisingly, we found an almost equal amount of spike protein inside and on the surface of bloodborne EVs. This would have proven difficult to determine using other conventional methods. Conclusion: To summarize, we have developed an easy-to-perform, sensitive, and highly efficient method that offers a mechanism to examine bloodborne EV cargo without disrupting their structural integrity.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135508237","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Receptor activator of nuclear factor-kappa B is enriched in CD9-positive extracellular vesicles released by osteoclasts. 核因子κB受体激活剂在破骨细胞释放的CD9阳性细胞外小泡中富集。

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 Epub Date: 2023-09-06 DOI: 10.20517/evcna.2023.38

Shaobo Ruan, Wellington J Rody, Shivani S Patel, Lina I Hammadi, Macey L Martin, Lorraine P de Faria, George Daaboul, Leif S Anderson, Mei He, L Shannon Holliday

{"title":"Receptor activator of nuclear factor-kappa B is enriched in CD9-positive extracellular vesicles released by osteoclasts.","authors":"Shaobo Ruan, Wellington J Rody, Shivani S Patel, Lina I Hammadi, Macey L Martin, Lorraine P de Faria, George Daaboul, Leif S Anderson, Mei He, L Shannon Holliday","doi":"10.20517/evcna.2023.38","DOIUrl":"10.20517/evcna.2023.38","url":null,"abstract":"Aim: Receptor activator of nuclear factor-kappa B (RANK)-containing extracellular vesicles (EVs) bind RANK-Ligand (RANKL) on osteoblasts, and thereby simultaneously inhibit bone resorption and promote bone formation. Because of this, they are attractive candidates for therapeutic bone anabolic agents. Previously, RANK was detected in 1 in every 36 EVs from osteoclasts by immunogold electron microscopy. Here, we have sought to characterize the subpopulation of EVs from osteoclasts that contains RANK in more detail.Methods: The tetraspanins CD9 and CD81 were localized in osteoclasts by immunofluorescence. EVs were visualized by transmission electron microscopy. A Single Particle Interferometric Reflectance Imaging Sensor (SP-IRIS) and immunoaffinity isolations examined whether RANK is enriched in specific types of EVs.Results: Immunofluorescence showed CD9 was mostly on or near the plasma membrane of osteoclasts. In contrast, CD81 was localized deeper in the osteoclast's cytosolic vesicular network. By interferometry, both CD9 and CD81 positive EVs from osteoclasts were small (56-83 nm in diameter), consistent with electron microscopy. The CD9 and CD81 EV populations were mostly distinct, and only 22% of the EVs contained both markers. RANK was detected by SP-IRIS in 2%-4% of the CD9-containing EVs, but not in CD81-positive EVs, from mature osteoclasts. Immunomagnetic isolation of CD9-containing EVs from conditioned media of osteoclasts removed most of the RANK. A trace amount of RANK was isolated with CD81.Conclusion: RANK was enriched in a subset of the CD9-positive EVs. The current study provides the first report of selective localization of RANK in subsets of EVs.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10629932/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"71489572","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

New insights of engineered extracellular vesicles as promising therapeutic systems 工程细胞外囊泡作为有前途的治疗系统的新见解

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2023.22

Cheng Jiang, Hongxing Liu, Yuhui Liao, Yanyan Jiang

引用次数: 0

Opportunities for engineering outer membrane vesicles using synthetic biology approaches 利用合成生物学方法设计外膜囊泡的机会

Extracellular vesicles and circulating nucleic acids Pub Date : 2023-01-01 DOI: 10.20517/evcna.2023.21

Richard J. R. Kelwick, A. Webb, P. Freemont

{"title":"Opportunities for engineering outer membrane vesicles using synthetic biology approaches","authors":"Richard J. R. Kelwick, A. Webb, P. Freemont","doi":"10.20517/evcna.2023.21","DOIUrl":"https://doi.org/10.20517/evcna.2023.21","url":null,"abstract":"Gram-negative bacteria naturally shed lipid vesicles, which contain complex molecular cargoes, from their outer membrane. These outer membrane vesicles (OMVs) have important biological functions relating to microbial stress responses, microbiome regulation, and host-pathogen interactions. OMVs are also attractive vehicles for delivering drugs, vaccines, and other therapeutic agents because of their ability to interact with host cells and their natural immunogenic properties. OMVs are also set to have a positive impact on other biotechnological and medical applications including diagnostics, bioremediation, and metabolic engineering. We envision that the field of synthetic biology offers a compelling opportunity to further expand and accelerate the foundational research and downstream applications of OMVs in a range of applications including the provision of OMV-based healthcare technologies. In our opinion, we discuss how current and potential future synergies between OMV research and synthetic biology approaches might help to further accelerate OMV research and real-world applications for the benefit of animal and human health.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2023-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"82802165","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A report on ASIC2021: a conference on extracellular vesicle communication mechanisms. ASIC2021报告:细胞外囊泡通讯机制会议

Extracellular vesicles and circulating nucleic acids Pub Date : 2022-01-01 DOI: 10.20517/evcna.2022.31

Ashley E Russell, Susmita Sil, Shilpa Buch, Michael W Graner

{"title":"A report on ASIC2021: a conference on extracellular vesicle communication mechanisms.","authors":"Ashley E Russell, Susmita Sil, Shilpa Buch, Michael W Graner","doi":"10.20517/evcna.2022.31","DOIUrl":"https://doi.org/10.20517/evcna.2022.31","url":null,"abstract":"AD. His group generated human induced pluripotent stem cells (hiPSCs) and differentiated them into neuronal, astrocytic, oligodendrocytic, and microglial cell types. Proteomic profiles of EVs from these differentiated iPSC cells contained cell-type specific markers: excitatory neurons (ATP1A3, NCAM1); astrocytes (LRP1, ITGA6); microglia-like cells (ITGAM, CD300A); and oligodendrocyte-like cells (LAMP2, FTH1). There were also 16 pan-EV marker candidates, including integrins and annexins. Cell type-specific EV proteins could also be found when comparing their data to CSF EV proteomic datasets, which also held true for brain-derived EVs. Correlation networks and pathway analyses identified proteins in each cell subset EVs with co-expression in AD. It was shown that astrocyte-specific EV (ADEV) markers were most significantly associated with AD pathology and cognitive impairment, thereby underscoring the role of ADEVs in AD progression. The hub protein from this module, integrin- β 1 (ITGB1), was elevated in ADEVs purified from total brain-derived EVs and associated with brain A β 42 and tau load in independent cohorts. From this, it was found that astrocytes are likely in an activated state due to IL1B, and astrocytic AD EVs are enriched in ITGB1. This correlated with A β 42 and phosphoTau, and these EVs enhance neuronal uptake via integrin signaling. Thus, this study provides a featured framework and rich resource for analyses of EV functions in neurodegenerative diseases in a cell type-specific manner.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10147373/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9768443","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

Towards responsible ctDNA-based multi-cancer screening: a preliminary exploration and discussion of ethically relevant aspects 面向负责任的基于ctdna的多癌筛查:伦理相关方面的初步探索与讨论

Extracellular vesicles and circulating nucleic acids Pub Date : 2022-01-01 DOI: 10.20517/evcna.2022.23

W. Dondorp

{"title":"Towards responsible ctDNA-based multi-cancer screening: a preliminary exploration and discussion of ethically relevant aspects","authors":"W. Dondorp","doi":"10.20517/evcna.2022.23","DOIUrl":"https://doi.org/10.20517/evcna.2022.23","url":null,"abstract":"While testing for easily accessible biomarkers in the circulation (“liquid biopsy”) has found its way to clinical cancer care, a further expected development is its use as a “universal” early detection test in population screening for cancer. A promising marker for such screening is circulating cell-free fragments of tumor DNA, shed into the circulation during tumor cell turnover. Several blood-based “multicancer early detection (MCED) tests” have recently been developed - but still need validation in large-scale studies involving non-patient populations. In this paper, we proactively explore the ethical aspects of this development. We refer to an often quoted synthesis of the internationally accepted framework of principles for responsible screening as first drawn up for the World Health Organisation (WHO) by Wilson and Junger 50 years ago and further developed and fine-tuned ever since. As our analysis suggests, some specific ethical issues and concerns about potential MCED screening connect to the fact that cancer is not just one disease. As a consequence, not all findings will have the same clinical utility. We discuss this against the background of earlier debates pertaining to broad scope forms of screening in other contexts, specifically newborn and reproductive genetic screening. We highlight the guidance provided by some of the criteria from the screening framework that seems most relevant in this connection: the need for screening objectives to be defined at the outset, the need for mechanisms to minimize potential risks, and the requirement that, for those participating in the screening, the overall benefits outweigh the harm.","PeriodicalId":73008,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2022-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"74777617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 1