scholarly journals Mechanisms of Extracellular Vesicle Biogenesis, Cargo Loading, and Release

2021 ◽  
Author(s):  
Abdel A. Alli
Keyword(s):  
Plasma Membrane ◽  
Circadian Rhythms ◽  
Nucleic Acids ◽  
Extracellular Vesicles ◽  
Extracellular Space ◽  
Extracellular Vesicle ◽  
Apoptotic Bodies ◽  
Endosomal Membranes ◽  
Vesicle Biogenesis

Extracellular vesicles (EVs) are carriers of various biomolecules including bioactive enzymes, lipids, proteins, nucleic acids, and metabolites. EVs are classified into three main types based on their size, biogenesis, and cargo. Exosomes originate from endosomal membranes and are the smallest type of EV. Microvesicles (MVs) or microparticles are larger in size, and like apoptotic bodies which represent the largest type of EVs, both of these vesicles originate from outward budding of the plasma membrane. As discussed in this chapter, cargo loading of EVs and their release into the extracellular space where they can be taken up by neighboring or distant cells plays an important role in physiology and pathophysiology. This chapter will outline specific mechanisms involved in the loading and enrichment of miRNAs, proteins, and lipids within EVs. As explained here, various external and biological stimuli play a role in EV release. Finally, recent studies have shown that the biogenesis, cargo loading, and release of EVs are governed by circadian rhythms. Although EVs were once thought to serve as garbage disposals of cells, the numerous roles they serve in physiology and pathophysiology are now being appreciated.

scholarly journals Evaluation of plasmatic extracellular vesicles by size

2021 ◽  
Author(s):  
Laura Cantone ◽  
Mirjam Hoxha ◽  
Chiara Favero ◽  
Luca Ferrari ◽  
Valentina Bollati
Keyword(s):  
Plasma Membrane ◽  
Blood Plasma ◽  
Extracellular Vesicles ◽  
High Precision ◽  
Extracellular Vesicle ◽  
Nanoparticle Tracking Analysis ◽  
Research Needs ◽  
Plasma Samples ◽  
Constant Flow ◽  
Syringe Pump

Abstract Extracellular vesicles (EVs) play a key role in many physiological and pathological processes [1]. EVs are a heterogeneous group of membrane-confined particles including endosome-derived exosomes and plasma membrane-originated microvesicles. The expanding field of extracellular vesicle research needs reproducible and accurate methods to characterize EVs [2]. EV profiling can be challenging due to the small size and heterogeneity. This protocol aims to provide a method to isolate EVs and facilitate high-precision particle quantitation by Nanoparticle Tracking Analysis (NTA)[3, 4]. NTA is commonly used to determine EV concentration and diameter [5, 6]. The protocol here described refers to the isolation of EVs from blood-plasma samples by using ultracentrifugation and then quantification and sizing of EVs with NTA by NanoSight NS300 system (Malvern Panalytical Ltd., Malvern, UK) provided with a syringe pump module enabling analysis in constant flow for improved sample statistics.

scholarly journals Effects of ATP9A on Extracellular Vesicle Release and Exosomal Lipid Composition

2020 ◽  
Vol 2020 ◽  
pp. 1-17
Author(s):  
Xiao Xu ◽  
Limei Xu ◽  
Peng Zhang ◽  
Kan Ouyang ◽  
Yin Xiao ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Extracellular Vesicles ◽  
Lipid Composition ◽  
Extracellular Vesicle ◽  
Biological Processes ◽  
Vesicle Release ◽  
Endosomal Recycling ◽  
Intracellular Compartments ◽  
Intercellular Communications

Numerous biological processes are regulated by the intercellular communications arising from extracellular vesicles (EVs) released from cells. However, the mechanisms that regulate the quantity of EV discharged have yet to be understood. While it is known that ATP9A, a P4-ATPase, is involved in endosomal recycling, it is not clear whether it also contributes to the release of EVs and the makeup of exosomal lipids. This study is aimed at exploring the role of human ATP9A in the process of EV release and, further, to analyze the profiles of EV lipids regulated by ATP9A. Our results demonstrate that ATP9A is located in both the intracellular compartments and the plasma membrane. The percentage of ceramides and sphingosine was found to be significantly greater in the control cells than in the ATP9A overexpression and ATP9A knockout groups. However, EV release was greater in ATP9A knockout cells, indicating that ATP9A inhibits the release of EVs. This study revealed the effects of ATP9A on the release of EVs and the lipid composition of exosomes.

Dynamin-like proteins are essential for vesicle biogenesis in Mycobacterium tuberculosis

2020 ◽  
Author(s):  
Shamba Gupta ◽  
Ainhoa Palacios ◽  
Atul Khataokar ◽  
Brian Weinrick ◽  
Jose L. Lavín ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Extracellular Vesicles ◽  
Cell Envelope ◽  
Membrane Vesicle ◽  
Extracellular Vesicle ◽  
Dependent Manner ◽  
Surface Lipid ◽  
Cell Infection ◽  
Vesicle Release ◽  
Vesicle Biogenesis

ABSTRACTMycobacterium tuberculosis (Mtb) secretes pathogenicity factors and immunologically active molecules via membrane vesicles. However, nothing is known about the mechanisms involved in mycobacterial vesicle biogenesis. This study investigates molecular determinants of membrane vesicle production in Mtb by analyzing Mtb cells under conditions of high vesicle production: iron limitation and VirR restriction. Ultrastructural analysis showed extensive cell envelope restructuring in association with vesicle release that correlated with downregulation of cell surface lipid biosynthesis and peptidoglycan alterations. Comparative transcriptomics showed common upregulation of the iniBAC operon in association with high vesicle production in Mtb cells. Vesicle production analysis demonstrated that the dynamin-like proteins (DLPs) encoded by this operon, IniA and IniC, are necessary for release of EV by Mtb in culture and in infected macrophages. Isoniazid, a first-line antibiotic, used in tuberculosis treatment, was found to stimulate vesicle release in a DLP-dependent manner. Our results provide a new understanding of the function of mycobacterial DLPs and mechanistic insights into vesicle biogenesis. The findings will enable further understanding of the relevance of Mtb-derived extracellular vesicles in the pathogenesis of tuberculosis and may open new avenues for therapeutic research.IMPORTANCEIron is an essential nutrient that promotes survival and growth of M. tuberculosis, the bacterium that causes human tuberculosis (TB). Limited availability of iron, often encountered in the host environment, stimulates M. tuberculosis to secrete membrane-bound extracellular vesicles containing molecules that may help it evade the immune system. Characterizing the bacterial factors and mechanisms involved in the production of mycobacterial vesicles is important for envisioning ways to interfere with this process. Here, we report the discovery of proteins required by M. tuberculosis for vesicle biogenesis in culture and during host cell infection. We also demonstrate a connection between antibiotic response and extracellular vesicle production. The work provides insights into the mechanisms underlying vesicle biogenesis in M. tuberculosis and permits better understanding of the significance of vesicle production to M. tuberculosis-host interactions and antibiotic stress response.

scholarly journals Alcohol Exposure Impacts the Composition of HeLa-Derived Extracellular Vesicles

Biomedicines ◽  
2019 ◽  
Vol 7 (4) ◽  
pp. 78 ◽  
Author(s):  
Leandra B. Jones ◽  
Sanjay Kumar ◽  
Aliyah J. Curry ◽  
Jayde S. Price ◽  
Alexandre Krendelchtchikov ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Hela Cells ◽  
Critical Role ◽  
Alcohol Exposure ◽  
Extracellular Vesicle ◽  
Exosome Biogenesis ◽  
Cancer Line ◽  
Vesicle Biogenesis ◽  
Progression Of Disease ◽  
Shock Proteins

Extracellular vesicles are nanosized vesicles that are under intense investigation for their role in intercellular communication. Extracellular vesicles have begun to be examined for their role in disease protection and their role as disease biomarkers and/or vaccine agents. The goal of this study was to investigate the effects of alcohol exposure on the biogenesis and composition of extracellular vesicles derived from the cervical cancer line, HeLa. The HeLa cells were cultured in exosome-free media and were either mock-treated (control) or treated with 50 mM or 100 mM of alcohol for 24 h and 48 h. Our results demonstrated that alcohol significantly impacts HeLa cell viability and exosome biogenesis/composition. Importantly, our studies demonstrate the critical role of alcohol on HeLa cells, as well as HeLa-derived extracellular vesicle biogenesis and composition. Specifically, these results indicate that alcohol alters extracellular vesicles’ packaging of heat shock proteins and apoptotic proteins. Extracellular vesicles serve as communicators for HeLa cells, as well as biomarkers for the initiation and progression of disease.

scholarly journals Quantitative Proteomic Analysis of Biogenesis-Based Classification for Extracellular Vesicles

Proteomes ◽  
2020 ◽  
Vol 8 (4) ◽  
pp. 33
Author(s):  
Linwen Zhang ◽  
Jeremie Parot ◽  
Vincent A. Hackley ◽  
Illarion V. Turko
Keyword(s):  
Mass Spectrometry ◽  
Plasma Membrane ◽  
Extracellular Vesicles ◽  
Specific Protein ◽  
Mass Spectrometry Analysis ◽  
Protein Markers ◽  
Spectrometry Analysis ◽  
Endosomal Membrane ◽  
Size Limitation ◽  
Endosomal Membranes

Extracellular vesicles (EVs) are traditionally divided into two major groups: (i) large vesicles originating from plasma membrane and called microvesicles, and (ii) small vesicles originating from the endoplasmic membrane and called exosomes. However, it is increasingly clear that the actual composition of a particular EV preparation cannot be adequately described with these two simple terms and is much more complex. Since the cell membrane origin of EVs predetermines their biological functions, the understanding of EV biogenesis is important for accurate interpretation of observed results. In the present study, we propose to take advantage of selective expression of some proteins in plasma or endosomal membranes and to use these proteins as plasma membrane-specific or endosomal membrane-specific markers. We have demonstrated that a quantitative mass spectrometry analysis allows simultaneous measurement of plasma membrane-specific and endosomal membrane-specific proteins in microvesicles and exosomes obtained after differential ultracentrifugation. Before mass spectrometry analysis, we also used sonicated platelets as a model of mixed EVs and multidetector asymmetrical-flow field-flow fractionation as an analytical method to verify a possible cross contamination of obtained microvesicles and exosomes. Based on the quantitative appearance of membrane-specific protein markers in EV preparations from human plasma and from human ARPE-19 cell medium, we concluded that there is no actual size limitation and both microvesicles and exosomes can be represented by large and small vesicles.

scholarly journals Extracellular vesicles: communication, coercion, and conditioning

2013 ◽  
Vol 24 (9) ◽  
pp. 1253-1259 ◽  
Author(s):  
David A. Shifrin ◽  
Michelle Demory Beckler ◽  
Robert J. Coffey ◽  
Matthew J. Tyska
Keyword(s):  
Extracellular Vesicles ◽  
Extracellular Space ◽  
Cell Biology ◽  
Biological Significance ◽  
Local Environment ◽  
Cell Types ◽  
Extracellular Vesicle ◽  
Functional Studies ◽  
Wide Range ◽  
Physiological Homeostasis

Cells communicate with neighboring cells and condition their local environment by secreting soluble factors into the extracellular space. These well-studied facets of cell biology are essential for the establishment and maintenance of physiological homeostasis. However, accumulating evidence has revealed that specific ligands, enzymes, and macromolecules are distributed into the extracellular space by virtue of their association with small vesicles, which are released by a variety of cell types. Although the biological significance of such vesicles was initially debated, purification and subsequent functional studies have shown that these extracellular vesicles are bioactive organelles carrying a wide range of protein and nucleic acid cargoes. In many cases these vesicles are laden with molecules that are involved in cell signaling, although other diverse functions are being revealed at a rapid pace. In this Perspective, we discuss recent developments in the understanding of the major pathways of extracellular vesicle biogenesis and how these vesicles contribute to the maintenance of physiological homeostasis.

Roles of glia-derived extracellular vesicles in central nervous system diseases: an update

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Hao Sun ◽  
Xiaojuan Su ◽  
Shiping Li ◽  
Dezhi Mu ◽  
Yi Qu
Keyword(s):  
Central Nervous System ◽  
Nervous System ◽  
Nucleic Acids ◽  
Extracellular Vesicles ◽  
Extracellular Space ◽  
Cell Communication ◽  
Heterogeneous Group ◽  
Nervous System Diseases ◽  
Central Nervous System Diseases ◽  
Cns Diseases

Abstract Extracellular vesicles (EVs) are a heterogeneous group of cell-derived membranous vesicles secreted by various cells in the extracellular space. Accumulating evidence shows that EVs regulate cell-to-cell communication and signaling in the pathological processes of various diseases by carrying proteins, lipids, and nucleic acids to recipient cells. Glia-derived EVs act as a double-edged sword in the pathogenesis of central nervous system (CNS) diseases. They may be vectors for the spread of diseases or act as effective clearance systems to protect tissues. In this review, we summarize recent studies on glia-derived EVs with a focus on their relationships with CNS diseases.

scholarly journals SMPD3-mediated extracellular vesicle biogenesis inhibits oligodendroglioma growth

2020 ◽  
Author(s):  
Anjali Balakrishnan ◽  
Lata Adnani ◽  
Vorapin Chinchalongporn ◽  
Lakshmy Vasan ◽  
Oleksandr Prokopchuk ◽  
...  
Keyword(s):  
Cell Lines ◽  
Extracellular Vesicles ◽  
Extracellular Vesicle ◽  
Driver Mutations ◽  
Lower Grade ◽  
Neoplastic Cells ◽  
Growth Inhibitory ◽  
Vesicle Biogenesis ◽  
Slow Growing

Abstract Oligodendrogliomas are lower-grade, slow-growing gliomas that are ultimately fatal. Although driver mutations are known, the mechanisms underlying their signature slow growth rates are poorly understood. We found evidence for intra-tumoral interactions between neoplastic and non-neoplastic cells in oligodendroglioma tissues. To further study these cell interactions, we used two patient-derived oligodendroglioma cell lines of lower and higher aggressivity. Both oligodendroglioma cell lines released extracellular vesicles that had cytotoxic effects on non-neoplastic and neoplastic cells, but each had distinct vesicular proteomes. Consistent with extracellular vesicles mediating growth inhibitory effects in oligodendrogliomas, higher expression levels of several extracellular vesicle biogenesis genes (SMPD3, TSG101, STAM1) correlates with longer survival in oligodendroglioma patients. Furthermore, SMPD3 overexpression slows oligodendroglioma cell growth in culture. Conversely, SMPD3 knockdown enhances oligodendroglioma proliferation in vitro, in murine xenografts, and in human cerebral organoid co-cultures. Oligodendroglioma-derived extracellular vesicles thus mediate tumor cell microenvironmental interactions that contribute to low aggressivity.

scholarly journals Tetraspanins, More than Markers of Extracellular Vesicles in Reproduction

2020 ◽  
Vol 21 (20) ◽  
pp. 7568
Author(s):  
Jana Jankovičová ◽  
Petra Sečová ◽  
Katarína Michalková ◽  
Jana Antalíková
Keyword(s):  
Extracellular Vesicles ◽  
Reproductive System ◽  
Extracellular Vesicle ◽  
Cell Uptake ◽  
Cell Contact ◽  
Cellular Processes ◽  
Pathological Conditions ◽  
Vesicle Biogenesis ◽  
Future Direction ◽  
Insight Into

The participation of extracellular vesicles in many cellular processes, including reproduction, is unquestionable. Although currently, the tetraspanin proteins found in extracellular vesicles are mostly applied as markers, increasing evidence points to their role in extracellular vesicle biogenesis, cargo selection, cell targeting, and cell uptake under both physiological and pathological conditions. In this review, we bring other insight into the involvement of tetraspanin proteins in extracellular vesicle physiology in mammalian reproduction. We provide knowledge regarding the involvement of extracellular vesicle tetraspanins in these processes in somatic cells. Furthermore, we discuss the future direction towards an understanding of their functions in the tissues and fluids of the mammalian reproductive system in gamete maturation, fertilization, and embryo development; their involvement in mutual cell contact and communication in their complexity.

scholarly journals Novel Epigenetic Biomarkers in Pregnancy-Related Disorders and Cancers

Cells ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 1459 ◽  
Author(s):  
Karin-Kujundzic ◽  
Sola ◽  
Predavec ◽  
Potkonjak ◽  
Somen ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Extracellular Vesicles ◽  
Diagnostic Tools ◽  
Epigenetic Changes ◽  
Apoptotic Bodies ◽  
Diagnostic Biomarkers ◽  
Epigenetic Biomarkers ◽  
Circulating Nucleic Acids ◽  
Specific Proteins ◽  
In Pregnancy

As the majority of cancers and gestational diseases are prognostically stage- and grade-dependent, the ultimate goal of ongoing studies in precision medicine is to provide early and timely diagnosis of such disorders. These studies have enabled the development of various new diagnostic biomarkers, such as free circulating nucleic acids, and detection of their epigenetic changes. Recently, extracellular vesicles including exosomes, microvesicles, oncosomes, and apoptotic bodies have been recognized as powerful diagnostic tools. Extracellular vesicles carry specific proteins, lipids, DNAs, mRNAs, and miRNAs of the cells that produced them, thus reflecting the function of these cells. It is believed that exosomes, in particular, may be the optimal biomarkers of pathological pregnancies and cancers, especially those that are frequently diagnosed at an advanced stage, such as ovarian cancer. In the present review, we survey and critically appraise novel epigenetic biomarkers related to free circulating nucleic acids and extracellular vesicles, focusing especially on their status in trophoblasts (pregnancy) and neoplastic cells (cancers).

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