Pevindu Abeysinghe, Natalie Turner, Murray D Mitchell
{"title":"A comparative analysis of small extracellular vesicle (sEV) micro-RNA (miRNA) isolation and sequencing procedures in blood plasma samples.","authors":"Pevindu Abeysinghe, Natalie Turner, Murray D Mitchell","doi":"10.20517/evcna.2023.55","DOIUrl":"10.20517/evcna.2023.55","url":null,"abstract":"<p><strong>Aims: </strong>Analysis of miRNA (18-23nt) encapsulated in small extracellular vesicles (sEVs) (diameter ~30-200 nm) is critical in understanding the diagnostic and therapeutic value of sEV miRNA. However, various sEV enrichment techniques yield different quantities and qualities of sEV miRNA. Here, we compare the efficacy of three sEV isolation techniques in four combinations for miRNA next-generation sequencing.</p><p><strong>Methods: </strong>Blood plasma from four Holstein-Friesian dairy cows (<i>Bos taurus</i>) (<i>n</i> = 4) with similar genetic traits and physical characteristics were pooled to isolate sEV. Ultracentrifugation (UC) (100,000 × <i>g</i>, 2 h at 4 °C), size-exclusion chromatography (SEC) and ultrafiltration (UF) were used to design four groups of sEV isolations (UC+SEC, SEC+UC, SEC+UF and UC+SEC+UF). sEV miRNAs were isolated using a combination of TRIzol, Chloroform and miRNeasy mini kit (<i>n</i> = 4/each), later sequenced utilizing Novaseq S1 platform (single-end 100 bp sequencing).</p><p><strong>Results: </strong>All four sEV methods yielded > 1,700 miRNAs and sEV miRNAs demonstrated a clear separation from control blood plasma circulating miRNA (PCA analysis). MiR-381-3p, miR-23-3p, and miR-18b-3p are among the 25 miRNAs unique to sEV, indicating potential sEV-specific miRNA markers. Further, those 25 miRNAs mostly regulate immune-related functions, indicating the value of sEV miRNA cargo in immunology.</p><p><strong>Conclusion: </strong>The four sEV miRNA isolation methods employed in this study are valid techniques. The choice of method depends on the research question and study design. If purity is of concern, the UC+SEC method resulted in the best particles/µg protein ratio, which is often used as an indication of sample purity. These results could eventually establish sEV miRNAs as effective diagnostic and therapeutic tools of immunology.</p>","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"5 1","pages":"119-137"},"PeriodicalIF":0.0,"publicationDate":"2024-02-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648519/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857479","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}
Naveed Akbar, Evelyn Grace Luciani, Raheel Ahmad, Dasol Lee, Sara Veiga, Daniel Christopher Rabe, Shannon Leigh Stott
{"title":"The isolation of VCAM-1<sup>+</sup> endothelial cell-derived extracellular vesicles using microfluidics.","authors":"Naveed Akbar, Evelyn Grace Luciani, Raheel Ahmad, Dasol Lee, Sara Veiga, Daniel Christopher Rabe, Shannon Leigh Stott","doi":"10.20517/evcna.2023.51","DOIUrl":"10.20517/evcna.2023.51","url":null,"abstract":"<p><p><b>Background:</b> Vascular cell adhesion molecule-1 (VCAM-1<sup>+</sup>) endothelial cell-derived extracellular vesicles (EC-EVs) are augmented in cardiovascular disease, where they can signal the deployment of immune cells from the splenic reserve. Endothelial cells in culture activated with pro-inflammatory tumor necrosis factor-α (TNF-a) also release VCAM-1<sup>+</sup> EC-EVs. However, isolating VCAM-1<sup>+</sup> EC-EVs from conditioned cell culture media for subsequent in-depth analysis remains challenging. <b>Aim:</b> We utilized the extracellular vesicles (EV) microfluidics herringbone chip (<sup>EV</sup>HB-Chip), coated with anti-VCAM-1 antibodies, for selective capture of VCAM-1<sup>+</sup> cells and EC-EVs. <b>Methods and Results:</b> Engineered EA.hy926 endothelial cells overexpressing VCAM-1 (<i>P</i> < 0.001 versus control) showed increased binding to the VCAM-1- <sup>EV</sup>HB-Chip versus an IgG device. TNF-α-stimulated human umbilical cord vein endothelial cells (HUVECs) exhibited elevated VCAM-1 protein levels (<i>P</i> < 0.001) and preferential binding to the VCAM-1- <sup>EV</sup>HB-Chip versus the IgG device. HUVECs stimulated with TNF-α showed differential gene expression of intercellular adhesion molecule-1 (ICAM-1) (<i>P</i> < 0.001) and VCAM-1 (<i>P</i> < 0.001) by digital droplet PCR versus control cells. HUVEC-derived EC-EVs were positive for CD9, CD63, HSP70, and ALIX and had a modal size of 83.5 nm. Control and TNF-α-stimulated HUVEC-derived EC-EV cultures were captured on the VCAM-1- <sup>EV</sup>HB-Chip, demonstrating selective capture. VCAM-1<sup>+</sup> EC-EV were significantly enriched for ICAM-1 (<i>P</i> < 0.001) mRNA transcripts. <b>Conclusion:</b> This study presents a novel approach using the <sup>EV</sup>HB-Chip, coated with anti-VCAM-1 antibodies and digital droplet PCR for the study of VCAM-1<sup>+</sup> EC-EVs. Isolation of VCAM-1<sup>+</sup> EC-EV from heterogeneous sources such as conditioned cell culture media holds promise for subsequent detailed characterization, and may facilitate the study of VCAM-1<sup>+</sup> EC-EVs in cardiovascular and metabolic diseases, for disease monitoring and therapeutic insights.</p>","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"5 1","pages":"83-94"},"PeriodicalIF":0.0,"publicationDate":"2024-02-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648473/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857533","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}
Xiaofang Zhang, Xiaofang Che, Sibo Zhang, Runze Wang, Mo Li, Yi Jin, Tianlu Wang, Yingqiu Song
{"title":"Mesenchymal stem cell-derived extracellular vesicles for human diseases.","authors":"Xiaofang Zhang, Xiaofang Che, Sibo Zhang, Runze Wang, Mo Li, Yi Jin, Tianlu Wang, Yingqiu Song","doi":"10.20517/evcna.2023.47","DOIUrl":"10.20517/evcna.2023.47","url":null,"abstract":"<p><p>Stem cell therapy is a novel approach for treating various severe and intractable diseases, including autoimmune disorders, organ transplants, tumors, and neurodegenerative diseases. Nevertheless, the extensive utilization of stem cells is constrained by potential tumorigenicity, challenges in precise differentiation, rejection concerns, and ethical considerations. Extracellular vesicles possess the ability to carry diverse bioactive factors from stem cells and deliver them to specific target cells or tissues. Moreover, they offer the advantage of low immunogenicity. Consequently, they have the potential to facilitate the therapeutic potential of stem cells, mitigating the risks associated with direct stem cell application. Therefore, the use of stem cell extracellular vesicles in clinical diseases has received increasing attention. This review summarizes advances in the use of extracellular vesicles from mesenchymal stem cells (MSC). MSC extracellular vesicles are used in the treatment of inflammatory diseases such as rheumatoid arthritis, liver injury, COVID-19, and allergies; in the repair of tissue damage in heart disease, kidney injury, and osteoarthritic diseases; as a carrier in the treatment of tumors; and as a regenerative agent in neurodegenerative disorders such as Alzheimer's and Parkinson's.</p>","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"5 1","pages":"64-82"},"PeriodicalIF":0.0,"publicationDate":"2024-02-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648454/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857532","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}
{"title":"Acknowledgment to reviewers of <i>Extracellular Vesicles and Circulating Nucleic Acids</i> in 2023.","authors":"Evcna Editorial Office","doi":"10.20517/evcna.2024.01","DOIUrl":"https://doi.org/10.20517/evcna.2024.01","url":null,"abstract":"","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"5 1","pages":"16-18"},"PeriodicalIF":0.0,"publicationDate":"2024-01-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648481/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857531","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}
Kumar Utkarsh, Namita Srivastava, Christopher Papayannakos, Ashima Nayyar, Azhar Khan, Shabirul Haque
{"title":"Breaking the silence: The role of extracellular vesicles in unraveling the diagnosis and treatment of endometriosis.","authors":"Kumar Utkarsh, Namita Srivastava, Christopher Papayannakos, Ashima Nayyar, Azhar Khan, Shabirul Haque","doi":"10.20517/evcna.2023.43","DOIUrl":"10.20517/evcna.2023.43","url":null,"abstract":"<p><p>Cell-to-cell communication is believed to be facilitated by membrane-bound vesicles called extracellular vesicles (EVs), which are released by cells. Protein, lipids, and nucleic acids are major cargo of EVs and are transported in these vesicles. Depending on the parent and recipient cell types, they can affect a wide variety of biological processes in the tissues to which they are delivered. EVs are essential for embryo implantation and endometriosis, and they are located in the uterine cavities of different species, where they promote blastocyst and endometrial preparation for implantation. This review focuses on what is currently understood regarding pathologic and diagnostic characteristics, and the potential therapeutic value of EVs in the context of endometriosis, where they can be used for drug delivery and targeted therapy due to their ability to carry bioactive molecules to specific cells or tissues. The findings of this review highlight the potential for a wide range of clinical applications that involve endometrial EVs in the areas of treatment, such as surgical and pharmacological, diagnostic biomarker development, and drug delivery systems, all with the ultimate goal of improving pregnancy success rates.</p>","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"4 4","pages":"599-614"},"PeriodicalIF":0.0,"publicationDate":"2023-12-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648450/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857471","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}
{"title":"Erratum: Identification of important extracellular vesicle RNA molecules related to sperm motility and prostate cancer.","authors":"Evcna Editorial Office","doi":"10.20517/evcna.2023.35","DOIUrl":"https://doi.org/10.20517/evcna.2023.35","url":null,"abstract":"<p><p>[This corrects the article DOI: 10.20517/evcna.2021.02.].</p>","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"4 3","pages":"338"},"PeriodicalIF":0.0,"publicationDate":"2023-07-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11651118/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857468","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}
{"title":"Acknowledgement to Reviewers of <i>Extracellular Vesicles and Circulating Nucleic Acids</i> in 2022.","authors":"Evcna Editorial Office","doi":"10.20517/evcna.2023.02","DOIUrl":"https://doi.org/10.20517/evcna.2023.02","url":null,"abstract":"","PeriodicalId":520322,"journal":{"name":"Extracellular vesicles and circulating nucleic acids","volume":"4 1","pages":"1-2"},"PeriodicalIF":0.0,"publicationDate":"2023-01-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11648434/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142857466","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}
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