Analyzing the Circulating MicroRNAs in Exosomes/Extracellular Vesicles from Serum or Plasma by qRT-PCR

Moldovan, Leni; Batte, Kara; Wang, Yijie; Wisler, Jon; Piper, Melissa G.

doi:10.1007/978-1-62703-453-1_10

Cited by 92 publications

(70 citation statements)

References 21 publications

(26 reference statements)

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“…The cargo can include genetic material such as mRNA, microRNA (miRNA), or even small amounts of DNA (Moldovan et al 2013), and proteins including transcription factors, cytokines, and growth factors, have also been described. Importantly, MVs also carry cellular receptors and transmembrane proteins on their surface characteristic of the cells from which they were released.…”

Section: Introductionmentioning

confidence: 99%

Microvesicles and exosomes: new players in metabolic and cardiovascular disease

Lawson¹,

Vicencio²,

Yellon³

et al. 2016

Journal of Endocrinology

285

241

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The past decade has witnessed an exponential increase in the number of publications referring to extracellular vesicles (EVs). For many years considered to be extracellular debris, EVs are now seen as novel mediators of endocrine signalling via cell-to-cell communication.With the capability of transferring proteins and nucleic acids from one cell to another, they have become an attractive focus of research for different pathological settings and are now regarded as both mediators and biomarkers of disease including cardio-metabolic disease. They also offer therapeutic potential as signalling agents capable of targeting tissues or cells with specific peptides or miRNAs. In this review, we focus on the role that microvesicles (MVs) and exosomes, the two most studied classes of EV, have in diabetes, cardiovascular disease, endothelial dysfunction, coagulopathies, and polycystic ovary syndrome. We also provide an overview of current developments in MV/exosome isolation techniques from plasma and other fluids, comparing different available commercial and non-commercial methods. We describe different techniques for their optical/biochemical characterization and quantitation. We also review the signalling pathways that exosomes and MVs activate in target cells and provide some insight into their use as biomarkers or potential therapeutic agents. In summary, we give an updated focus on the role that these exciting novel nanoparticles offer for the endocrine community.

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Section: Introductionmentioning

confidence: 99%

Microvesicles and exosomes: new players in metabolic and cardiovascular disease

Lawson¹,

Vicencio²,

Yellon³

et al. 2016

Journal of Endocrinology

285

241

View full text Add to dashboard Cite

show abstract

“…miRNAs are small, non-coding RNAs that have the ability to regulate whole networks of genes through transcriptional and translational regulation. In the circulation they are commonly found enclosed in extracellular vesicles (EVs) [7], bound to lipoproteins [8], or complexed with Argonaute-2 [9]. Furthermore, in vitro studies have shown that numerous cell types are capable of releasing miRNA in these complexes.…”

Section: Introductionmentioning

confidence: 99%

High Throughput Sequencing of Extracellular RNA from Human Plasma

et al. 2017

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The presence and relative stability of extracellular RNAs (exRNAs) in biofluids has led to an emerging recognition of their promise as ‘liquid biopsies’ for diseases. Most prior studies on discovery of exRNAs as disease-specific biomarkers have focused on microRNAs (miRNAs) using technologies such as qRT-PCR and microarrays. The recent application of next-generation sequencing to discovery of exRNA biomarkers has revealed the presence of potential novel miRNAs as well as other RNA species such as tRNAs, snoRNAs, piRNAs and lncRNAs in biofluids. At the same time, the use of RNA sequencing for biofluids poses unique challenges, including low amounts of input RNAs, the presence of exRNAs in different compartments with varying degrees of vulnerability to isolation techniques, and the high abundance of specific RNA species (thereby limiting the sensitivity of detection of less abundant species). Moreover, discovery in human diseases often relies on archival biospecimens of varying age and limiting amounts of samples. In this study, we have tested RNA isolation methods to optimize profiling exRNAs by RNA sequencing in individuals without any known diseases. Our findings are consistent with other recent studies that detect microRNAs and ribosomal RNAs as the major exRNA species in plasma. Similar to other recent studies, we found that the landscape of biofluid microRNA transcriptome is dominated by several abundant microRNAs that appear to comprise conserved extracellular miRNAs. There is reasonable correlation of sets of conserved miRNAs across biological replicates, and even across other data sets obtained at different investigative sites. Conversely, the detection of less abundant miRNAs is far more dependent on the exact methodology of RNA isolation and profiling. This study highlights the challenges in detecting and quantifying less abundant plasma miRNAs in health and disease using RNA sequencing platforms.

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“…In addition, the use of heparinase significantly increases the cost of RNA measurement. Recent studies also suggest that citrate treated plasma shows lower miRNA concentrations based on qPCR measurement compared to EDTA treated plasma from the same individual [56, 96]. …”

Section: Factors Affect the Spectrum Of Extracellular Rnamentioning

confidence: 99%

The Importance of Standardization on Analyzing Circulating RNA

et al. 2016

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Circulating RNAs, especially microRNAs (miRNAs) have recently emerged as non-invasive disease biomarkers. Despite enthusiasm and numerous reports on disease associated circulating miRNAs, currently there is no circulating miRNA-based diagnostic in use. In addition, there are many contradictory reports on the concentration changes of specific miRNA in circulation. Here we review the impact of various technical and non-technical factors related to circulating miRNA measurement and elucidate the importance of having a general guideline for sample preparation and concentration measurement in studying circulating RNA.

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Analyzing the Circulating MicroRNAs in Exosomes/Extracellular Vesicles from Serum or Plasma by qRT-PCR

Cited by 92 publications

References 21 publications

Microvesicles and exosomes: new players in metabolic and cardiovascular disease

Microvesicles and exosomes: new players in metabolic and cardiovascular disease

High Throughput Sequencing of Extracellular RNA from Human Plasma

The Importance of Standardization on Analyzing Circulating RNA

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