Comparison of methods of isolating extracellular vesicle microRNA from HepG2 cells for High-throughput sequencing

Xiong, Ying-Hui; Fan, Xue‐Gong; Chen, Yayu; Huang, Yan; Quan, Jun; Yi, Panpan; Xiao, Meifang; Huang, Zebing; Hu, Xingwang

doi:10.3389/fmolb.2022.976528

Cited by 3 publications

(3 citation statements)

References 29 publications

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“…Protein concentrations were detected with BCA Protein Assay according to the manufacturer’s protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies 52 , 53 .…”

Section: Methodsmentioning

confidence: 99%

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Xiao,

Huang,

Luo

et al. 2023

Sci Rep

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The coronavirus disease 2019 (COVID-19), which affects multiple organs, is causing an unprecedented global public health crisis. Most COVID-19 patients recover gradually upon appropriate interventions. Viruses were reported to utilize the small extracellular vesicles (sEVs), containing a cell-specific cargo of proteins, lipids, and nucleic acids, to escape the attack from the host’s immune system. This study aimed to examine the sEVs lipid profile of plasma of recovered COVID-19 patients (RCs). Plasma sEVs were separated from 83 RCs 3 months after discharge without underlying diseases, including 18 recovered asymptomatic patients (RAs), 32 recovered moderate patients (RMs), and 33 recovered severe and critical patients (RSs), and 19 healthy controls (HCs) by Total Exosome Isolation Kit. Lipids were extracted from sEVs and then subjected to targeted liquid chromatography-mass spectrometry. The size, concentration, and distribution of sEVs did not differ in RCs and HCs as validated by transmission electron microscopy, nanoparticle tracking analysis, and immunoblot analysis. Fifteen subclasses of 508 lipids were detected in plasma sEVs from HCs, RAs, RMs, and RSs, such as phosphatidylcholines (PCs) and diacylglycerols (DAGs), etc. Total lipid intensity displayed downregulation in RCs compared with HCs. The relative abundance of DAGs gradually dropped, whereas PCs, lysophosphatidylcholines, and sphingomyelins were higher in RCs relative to HCs, especially in RSs. 88 lipids out of 241 in sEVs of RCs were significantly different and a conspicuous increase was revealed with disease status. The sEVs lipids alternations were found to be significantly correlated with the clinical indices in RCs and HCs, suggesting that the impact of COVID-19 on lipid metabolism lingered for a long time. The lipid abnormalities bore an intimate link with glycerophospholipid metabolism and glycosylphosphatidylinositol anchor biosynthesis. Furthermore, the lipidomic analysis showed that RCs were at higher risk of developing diabetes and sustaining hepatic impairment. The abnormality of immunomodulation in RCs might still exist. The study may offer new insights into the mechanism of organ dysfunction and help identify novel therapeutic targets in the RCs.

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Section: Methodsmentioning

confidence: 99%

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Xiao,

Huang,

Luo

et al. 2023

Sci Rep

View full text Add to dashboard Cite

show abstract

“…LPC, a potent proin ammatory mediator, was increased in several types of acute lung injuries 41 . FFAs could enter mitochondria for beta-oxidation to produce ATP for viral replication and transmission, resulting in an impaired antiviral response (47). In COVID-19 acute patients, a concomitant increase in FFAs, such as FFA(18:1) and FFA(18:2), was observed (25).…”

Section: Plasma Lipids Alternations Linked With Covid-19 Pathogenesis...mentioning

confidence: 99%

“…Protein concentrations were detected with BCA Protein Assay according to the manufacturer's protocols. sEVs markers CD9 (Abcam, ab92726), CD63 (Abcam, ab193349), TSG101 (Abcam, ab83), and sEVs non-markers calnexin (Abcam, ab22595), GM130 (Abcam, ab52649), Lamin B1 (Abcam, ab16048), and GRP94 (Proteintech, 14700-1-AP) were measured to validate the expressions of sEVs as described in prior studies 46,47 .…”

Section: Western Blottingmentioning

confidence: 99%

Lipid metabolism of plasma-derived small extracellular in COVID-19 convalescent patients

Xiao

Huang

Luo

et al. 2023

Preprint

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The coronavirus disease 2019 (COVID-19), which affects multiple organs, is causing an unprecedented global public health crisis. Most COVID-19 patients recover gradually upon appropriate interventions. Viruses were reported to utilize the small extracellular vesicles (sEVs) to escape the attack from the host’s immune system. This study aimed to examine the lipid profile of plasma small extracellular vesicles of recovered COVID-19 patients (RCs). Plasma sEVs were separated from 83 RCs 3 months after discharge without underlying diseases, including 18 recovered asymptomatic patients (RAs), 32 recovered moderate patients (RMs), and 33 recovered severe and critical patients (RSs), and 19 healthy controls (HCs) by Total Exosome Isolation. Lipids were extracted from sEVs and then subjected to targeted liquid chromatography-mass spectrometry. Size, concentration, and distribution of plasma-derived sEVs from RAs, RMs, RSs, and HCs did not differ in RCs and HCs as validated by transmission electron microscopy, nanoparticle tracking analysis, and immunoblot analysis. Fifteen subclasses of 508 lipids were detected in plasma sEVs from HCs, RAs, RMs, and RSs, such as phosphatidylcholines (PCs) and diacylglycerols (DAGs), etc. Total lipid intensity displayed downregulation in RCs compared with HCs. The relative abundance of DAGs gradually dropped, whereas PCs, lysophosphatidylcholines, and sphingomyelins were higher in RCs relative to HCs, especially RSs. 88 lipids out of 241 were significantly different and a conspicuous increase in lipid profiles of RCs was revealed with disease status. The lipids alternations were found to be significantly correlated with the clinical indices in RCs and HCs, suggesting that the impact of COVID-19 on lipid metabolism lingered for a long time. The lipid abnormalities bore an intimate link with glycerophospholipid metabolism and glycosylphosphatidylinositol anchor biosynthesis. Furthermore, the lipidomic analysis showed that RCs were at higher risk of developing diabetes and sustaining hepatic impairment. The abnormality of immunomodulation in RCs might still exist. The study may offer new insights into the mechanism of organ dysfunction and help identify novel therapeutic targets in the RCs.

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Aptamer‐Functionalized Magnetic Ti₃C₂ Based Nanoplatform for Simultaneous Enrichment and Detection of Exosomes

Cui,

Zheng,

Qian

et al. 2024

Small

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Exosomes are nanovesicles secreted by cells, which play a crucial role in various pathological processes. Exosomes have shown great promise as tumor biomarkers because of the abundant secretion during tumor formation. The development of a convenient, efficient, and cost‐effective method for simultaneously enriching and detecting exosomes is of utmost importance for both basic research and clinical applications. In this study, an aptamer‐functionalized magnetic Ti3C2 composite material (Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM‐ssDNA) is prepared for the simultaneous enrichment and detection of exosomes. CD63 aptamers are utilized to recognize and capture the exosomes, followed by magnetic separation. The exosomes are then released by cleaving the disulfide bonds of DSP. Compared to traditional methods, Fe3O4@Ti3C2@PEI@DSP@aptamer@FAM‐ssDNA exhibited superior efficiency in enriching exosomes while preserving their structural and functional integrity. Detection of exosome concentration is achieved through the fluorescence quenching of Ti3C2 and the competitive binding between the exosomes and a fluorescently labeled probe. This method exhibited a low detection limit of 4.21 × 104 particles mL−1, a number that is comparable to the state‐of‐the‐art method in the detection of exosomes. The present study demonstrates a method of simultaneous enrichment and detection of exosomes with a high sensitivity, accuracy, specificity, and cost‐effectiveness providing significant potential for clinical research and diagnosis.

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Comparison of methods of isolating extracellular vesicle microRNA from HepG2 cells for High-throughput sequencing

Cited by 3 publications

References 29 publications

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Lipid metabolism of plasma-derived small extracellular in COVID-19 convalescent patients

Aptamer‐Functionalized Magnetic Ti₃C₂ Based Nanoplatform for Simultaneous Enrichment and Detection of Exosomes

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Comparison of methods of isolating extracellular vesicle microRNA from HepG2 cells for High-throughput sequencing

Cited by 3 publications

References 29 publications

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Lipid metabolism of plasma-derived small extracellular vesicles in COVID-19 convalescent patients

Lipid metabolism of plasma-derived small extracellular in COVID-19 convalescent patients

Aptamer‐Functionalized Magnetic Ti3C2 Based Nanoplatform for Simultaneous Enrichment and Detection of Exosomes

Contact Info

Product

Resources

About

Aptamer‐Functionalized Magnetic Ti₃C₂ Based Nanoplatform for Simultaneous Enrichment and Detection of Exosomes