No Significant Difference between Plasma miRNAs and Plasma-Derived Exosomal miRNAs from Healthy People

Tian, Fei; Shen, Yanting; Chen, Zhenzhu; Li, Rui; Ge, Qinyu

doi:10.1155/2017/1304816

Cited by 29 publications

(26 citation statements)

References 35 publications

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“…Taken together, our data show that a precipitation‐based EV isolation method removes only part of the vesicle‐free miRNAs, which has implications for interpretating previously published data. For example, a recent comparison of the miRNA contents of plasma and precipitation‐isolated EV pellets using a miRNA array indicated that the miRNA profile was comparable between the two preparations [33]. On the other hand, another report showed that EVs carry only a minor proportion of the plasma miRNAs [11].…”

Section: Discussionmentioning

confidence: 99%

Precipitation‐based extracellular vesicle isolation from rat plasma co‐precipitate vesicle‐free microRNAs

Karttunen

Heiskanen

Navarro-Ferrandis

et al. 2018

J of Extracellular Vesicle

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The microRNA (miRNA) cargo contained in plasma extracellular vesicles (EVs) offers a relatively little explored source of biomarkers for brain diseases that can be obtained noninvasively. Methods to isolate EVs from plasma, however, are still being developed. For EV isolation, it is important to ensure the removal of vesicle-free miRNAs, which account for approximately two-thirds of plasma miRNAs. Membrane particle precipitation-based EV isolation is an appealing method because of the simple protocol and high yield. Here, we evaluated the performance of a precipitation-based method to obtain enriched EV-specific miRNAs from a small volume of rat plasma. We performed size-exclusion chromatography (SEC) on precipitation-isolated EV pellets and whole plasma. The SEC fractions were analysed using Nanoparticle Tracking Analysis (NTA), protein and miRNA concentration assays, and droplet digital polymerase chain reaction for four miRNAs (miR-142-3p, miR-124-3p, miR-23a, miR-122). Precipitation-isolated EVs and selected SEC fractions from the plasma were also analysed with transmission electron microscopy (TEM). Precipitation-based EV isolation co-precipitated 9% to 15% of plasma proteins and 21% to 99% of vesicle-free miRNAs, depending on the individual miRNAs. In addition, the amount of miR-142-3p, found mainly in EV fractions, was decreased in the EV fractions, indicating that part of it was lost during precipitation-based isolation. Western blot and TEM revealed both protein and lipoprotein contamination in the precipitation-isolated EV-pellets. Our findings indicate that a precipitation-based method is not sufficient for purifying plasma EV-contained miRNA cargo. The particle number measured by NTA is high, but this is mostly due to the contaminating lipoproteins. Although a part of the vesicle-free miRNA is removed, vesicle-free miRNA still dominates in plasma EV pellets isolated by the precipitation-based method.

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Section: Discussionmentioning

confidence: 99%

Precipitation‐based extracellular vesicle isolation from rat plasma co‐precipitate vesicle‐free microRNAs

Karttunen

Heiskanen

Navarro-Ferrandis

et al. 2018

J of Extracellular Vesicle

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“…Several miRNAs were reported to differently express in plasma, serum, and peripheral blood exosomes. For example, miR-181b-5p and miR-21-5p were enriched in the exosomes of lung cancer patients instead of healthy individuals [ 86 ]. Plasma levels of miR-19b-3p, miR-21-5p, miR-221-3p, miR-409-3p, miR-425-5p, and miR-584-5p were elevated in lung adenocarcinoma patients, but only miR-19-3p, miR-21-5p, and miR-221-3p were found to be upregulated in plasma exosomes [ 84 ].…”

Section: The Disadvantage and Limitation Of Circulating Mirnas As Diamentioning

confidence: 99%

Circulating microRNAs as potential cancer biomarkers: the advantage and disadvantage

et al. 2018

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MicroRNAs are endogenous single-stranded non-coding small RNA molecules that can be secreted into the circulation and exist stably. They usually exhibit aberrant expression under different physiological and pathological conditions. Recently, differentially expressed circulating microRNAs were focused on as potential biomarkers for cancer screening. We herein review the role of circulating microRNAs for cancer diagnosis, tumor subtype classification, chemo- or radio-resistance monitoring, and outcome prognosis. Moreover, circulating microRNAs still have several issues hindering their reliability for the practical clinical application. Future studies need to elucidate further potential application of circulating microRNAs as specific and sensitive markers for clinical diagnosis or prognosis in cancers.

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“…Such a high number of sncRNAs with changed levels in MDS (almost 10-fold increase) suggests that mechanisms of sncRNA export into the blood circulation may be specifically affected in MDS, changing sncRNA profiles as a whole. Other studies also found no significant difference between plasma and exosomal miRNAs from healthy people [36]. On the other hand, differential regulation of miRNA levels between plasma and exosomes in different disease animal models has been described as well [37].…”

Section: Discussionmentioning

confidence: 96%

Circulating Small Noncoding RNAs Have Specific Expression Patterns in Plasma and Extracellular Vesicles in Myelodysplastic Syndromes and Are Predictive of Patient Outcome

Hruštincová

Krejčík

Kundrat

et al. 2020

Cells

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Myelodysplastic syndromes (MDS) are hematopoietic stem cell disorders with large heterogeneity at the clinical and molecular levels. As diagnostic procedures shift from bone marrow biopsies towards less invasive techniques, circulating small noncoding RNAs (sncRNAs) have become of particular interest as potential novel noninvasive biomarkers of the disease. We aimed to characterize the expression profiles of circulating sncRNAs of MDS patients and to search for specific RNAs applicable as potential biomarkers. We performed small RNA-seq in paired samples of total plasma and plasma-derived extracellular vesicles (EVs) obtained from 42 patients and 17 healthy controls and analyzed the data with respect to the stage of the disease, patient survival, response to azacitidine, mutational status, and RNA editing. Significantly higher amounts of RNA material and a striking imbalance in RNA content between plasma and EVs (more than 400 significantly deregulated sncRNAs) were found in MDS patients compared to healthy controls. Moreover, the RNA content of EV cargo was more homogeneous than that of total plasma, and different RNAs were deregulated in these two types of material. Differential expression analyses identified that many hematopoiesis-related miRNAs (e.g., miR-34a, miR-125a, and miR-150) were significantly increased in MDS and that miRNAs clustered on 14q32 were specifically increased in early MDS. Only low numbers of circulating sncRNAs were significantly associated with somatic mutations in the SF3B1 or DNMT3A genes. Survival analysis defined a signature of four sncRNAs (miR-1237-3p, U33, hsa_piR_019420, and miR-548av-5p measured in EVs) as the most significantly associated with overall survival (HR = 5.866, p < 0.001). In total plasma, we identified five circulating miRNAs (miR-423-5p, miR-126-3p, miR-151a-3p, miR-125a-5p, and miR-199a-3p) whose combined expression levels could predict the response to azacitidine treatment. In conclusion, our data demonstrate that circulating sncRNAs show specific patterns in MDS and that their expression changes during disease progression, providing a rationale for the potential clinical usefulness of circulating sncRNAs in MDS prognosis. However, monitoring sncRNA levels in total plasma or in the EV fraction does not reflect one another, instead, they seem to represent distinctive snapshots of the disease and the data should be interpreted circumspectly with respect to the type of material analyzed.

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No Significant Difference between Plasma miRNAs and Plasma-Derived Exosomal miRNAs from Healthy People

Cited by 29 publications

References 35 publications

Precipitation‐based extracellular vesicle isolation from rat plasma co‐precipitate vesicle‐free microRNAs

Precipitation‐based extracellular vesicle isolation from rat plasma co‐precipitate vesicle‐free microRNAs

Circulating microRNAs as potential cancer biomarkers: the advantage and disadvantage

Circulating Small Noncoding RNAs Have Specific Expression Patterns in Plasma and Extracellular Vesicles in Myelodysplastic Syndromes and Are Predictive of Patient Outcome

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