Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells

Verta, Roberta; Grange, Cristina; Škovroňová, Renata; Tanzi, Adele; Peruzzi, Licia; Deregibus, Maria Chiara; Camussi, Giovanni; Bussolati, Benedetta

doi:10.3390/cells11010146

Cited by 12 publications

(12 citation statements)

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“…Although there are few publications describing the characterization of HIV/AIDS exosome samples using this platform, it has been used to characterize EVs in the context of other viral infectious diseases such as herpes simplex [ 276 ] and COVID‐19. [ 277 ] This platform was used to characterize EVs from herpes simplex 1 virus (HSV‐1)‐infected patients, by measuring levels of the tetraspanins CD63, CD81, and CD9. [ 276 ] The platform was also used to characterize EV CD9, CD63, and CD81 to support the development of therapeutic approaches for blocking SARS‐CoV‐2 cell binding.…”

Section: Exosome Characterizationmentioning

confidence: 99%

“…[ 276 ] The platform was also used to characterize EV CD9, CD63, and CD81 to support the development of therapeutic approaches for blocking SARS‐CoV‐2 cell binding. [ 277 ] The use of the ExoView R100 platform and other novel integrated miniature devices, such as microfluidic systems with plasmonic and electrochemical sensors to analyze exosomes for disease detection and therapeutics research, [ 276 , 277 , 278 ] can minimize the transfer of samples between instruments when multiple exosome characteristics are studied.…”

Section: Exosome Characterizationmentioning

confidence: 99%

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Exosome Processing and Characterization Approaches for Research and Technology Development

et al. 2022

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Exosomes are extracellular vesicles that share components of their parent cells and are attractive in biotechnology and biomedical research as potential disease biomarkers as well as therapeutic agents. Crucial to realizing this potential is the ability to manufacture high‐quality exosomes; however, unlike biologics such as proteins, exosomes lack standardized Good Manufacturing Practices for their processing and characterization. Furthermore, there is a lack of well‐characterized reference exosome materials to aid in selection of methods for exosome isolation, purification, and analysis. This review informs exosome research and technology development by comparing exosome processing and characterization methods and recommending exosome workflows. This review also provides a detailed introduction to exosomes, including their physical and chemical properties, roles in normal biological processes and in disease progression, and summarizes some of the on‐going clinical trials.

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Section: Exosome Characterizationmentioning

confidence: 99%

Section: Exosome Characterizationmentioning

confidence: 99%

Exosome Processing and Characterization Approaches for Research and Technology Development

et al. 2022

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“…[49] Following this, many EV researchers performed dSTORMbased imaging of sEVs by labeling the EV canonical markers such as CD9, CD63, CD81, and TSG101 using the corresponding antibodies conjugated with different fluorophores. [50][51][52][53][54][55] Recently, STORM-based imaging was performed to reveal the heterogeneity in EV secretion by human macrophages. [56] The details of all the published studies based on SMLM for EV and their cargo characterization are furnished in Table 1.…”

Section: Smlm For Sev Imagingmentioning

confidence: 99%

Single Molecule Localization Microscopy for Studying Small Extracellular Vesicles

Ghanam

Chetty

Zhu

et al. 2023

Small

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guidelines suggests that EV researchers use the term small EVs (sEVs) for vesicles that are less than 200 nm in diameter. [4] sEVs contain many essential cargo biomolecules such as nucleic acids (DNA, mRNA, microRNA), proteins, and lipids. [1,3,5] sEVs have emerged as a functional mediator for communications between cells in health and disease. [1,6] sEVs derived from tumor cells contain disease-specific proteins, RNA, and double-stranded DNA (dsDNA), thereby representing the disease state and progression. [1,2,[7][8][9] Especially in tumor microenvironment, sEVs transfer their cargo from the tumor to stromal cells. [10,11] sEVs play pathophysiological roles in many other diseases, including neurodegenerative diseases, and various infections. They transmit important biomolecules that regulate many biological processes and influence the immune system. [1,[12][13][14][15] In the last decade, studies dealing with sEV biogenesis pathways and the role of sEVs in health and disease have grown exponentially. However, exploring the released sEVs and interaction of their cargo with cellular biomolecules in the distant recipient cells is significantly hampered by the lack of microscopy studies using improved imaging technology. Most of the current microscopy studies involving sEVs use conventional confocal microscopy to generate either 2D or 3D reconstruction images using Imaris. [11,[16][17][18][19][20][21][22] Although these images provide the information Small extracellular vesicles (sEVs) are 30-200 nm nanovesicles enriched with unique cargoes of nucleic acids, lipids, and proteins. sEVs are released by all cell types and have emerged as a critical mediator of cell-to-cell communication. Although many studies have dealt with the role of sEVs in health and disease, the exact mechanism of sEVs biogenesis and uptake remain unexplored due to the lack of suitable imaging technologies. For sEVs functional studies, imaging has long relied on conventional fluorescence microscopy that has only 200-300 nm resolution, thereby generating blurred images. To break this resolution limit, recent developments in super-resolution microscopy techniques, specifically single-molecule localization microscopy (SMLM), expanded the understanding of subcellular details at the few nanometer level. SMLM success relies on the use of appropriate fluorophores with excellent blinking properties. In this review, the basic principle of SMLM is highlighted and the state of the art of SMLM use in sEV biology is summarized. Next, how SMLM techniques implemented for cell imaging can be translated to sEV imaging is discussed by applying different labeling strategies to study sEV biogenesis and their biomolecular interaction with the distant recipient cells.

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“…EVs are ‘pass’ communicators to all parts of the organism and may be forced to carry SARS‐CoV‐2 viral fragments for further distant spread as the ‘prisoners’ (Elrashdy et al., 2020; Elrashdy et al., 2021; Pesce et al., 2022; Pironti et al., 2021) (Figure 5A). For example, SARS‐CoV‐2 RNA fragments can be detected in EVs (Ning et al., 2021), and EVs containing S2 or S1 proteins are more readily taken up by ACE2‐expressing cells (Verta et al., 2022). SARS‐CoV‐2 spike‐derived fragments are present in the exosomes of COVID‐19 patients, but are particularly enriched in mild COVID‐19 patients, and these exosomes can be used as a source of antigen presentation to enhance immune responses and through promote T‐cell activation (Pesce et al., 2022), whereas exosomes from severe COVID‐19 patients may lead to further deterioration of the COVID‐19 illness.…”

Section: Antagonism: Evs Antagonism With Sars‐cov‐2 Inspires Diagnosi...mentioning

confidence: 99%

Meet changes with constancy: Defence, antagonism, recovery, and immunity roles of extracellular vesicles in confronting SARS‐CoV‐2

Chen

Song

et al. 2022

J of Extracellular Vesicle

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Coronavirus disease 2019 (COVID‐19), caused by severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2), has wrought havoc on the world economy and people's daily lives. The inability to comprehensively control COVID‐19 is due to the difficulty of early and timely diagnosis, the lack of effective therapeutic drugs, and the limited effectiveness of vaccines. The body contains billions of extracellular vesicles (EVs), which have shown remarkable potential in disease diagnosis, drug development, and vaccine carriers. Recently, increasing evidence has indicated that EVs may participate or assist the body in defence, antagonism, recovery and acquired immunity against SARS‐CoV‐2. On the one hand, intercepting and decrypting the general intelligence carried in circulating EVs from COVID‐19 patients will provide an important hint for diagnosis and treatment; on the other hand, engineered EVs modified by gene editing in the laboratory will amplify the effectiveness of inhibiting infection, replication and destruction of ever‐mutating SARS‐CoV‐2, facilitating tissue repair and making a better vaccine. To comprehensively understand the interaction between EVs and SARS‐CoV‐2, providing new insights to overcome some difficulties in the diagnosis, prevention and treatment of COVID‐19, we conducted a rounded review in this area. We also explain numerous critical challenges that these tactics face before they enter the clinic, and this work will provide previous ‘meet change with constancy’ lessons for responding to future similar public health disasters. Extracellular vesicles (EVs) provide a ‘meet changes with constancy’ strategy to combat SARS‐CoV‐2 that spans defence, antagonism, recovery, and acquired immunity. Targets for COVID‐19 diagnosis, therapy, and prevention of progression may be found by capture of the message decoding in circulating EVs. Engineered and biomimetic EVs can boost effects of the natural EVs, especially anti‐SARS‐CoV‐2, targeted repair of damaged tissue, and improvement of vaccine efficacy.

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Generation of Spike-Extracellular Vesicles (S-EVs) as a Tool to Mimic SARS-CoV-2 Interaction with Host Cells

Cited by 12 publications

References 50 publications

Exosome Processing and Characterization Approaches for Research and Technology Development

Exosome Processing and Characterization Approaches for Research and Technology Development

Single Molecule Localization Microscopy for Studying Small Extracellular Vesicles

Meet changes with constancy: Defence, antagonism, recovery, and immunity roles of extracellular vesicles in confronting SARS‐CoV‐2

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