Diagnostic and Prognostic Potential of Extracellular Vesicles in Peripheral Blood

Revenfeld, Anne Louise; Bæk, Rikke; Nielsen, Mads; Stensballe, Allan; Varming, Kim; Jørgensen, Maléne Møller

doi:10.1016/j.clinthera.2014.05.008

Cited by 225 publications

(187 citation statements)

References 122 publications

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“…The contents of EV subsets present in patient biofluids, including serum, urine, semen, saliva, and bronchoalveolar lavage fluid, easily could be used as noninvasive biomarkers or indicators of the host's immune status (99). For example, it was determined via flow cytometry that sera from patients with chronic hepatitis C have elevated levels of EVs from CD4 + and CD8 + T cells, whereas patients with nonalcoholic fatty liver or nonalcoholic steatohepatitis have elevated levels of EVs from iNKT cells and CD14 + macrophages/monocytes.…”

Section: The Role Of Evs In Autoimmunitymentioning

confidence: 99%

Regulation of chronic inflammatory and immune processes by extracellular vesicles

Robbins

Dorronsoro

Booker

2016

Journal of Clinical Investigation

236

195

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Section: The Role Of Evs In Autoimmunitymentioning

confidence: 99%

Regulation of chronic inflammatory and immune processes by extracellular vesicles

Robbins

Dorronsoro

Booker

2016

Journal of Clinical Investigation

236

195

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“…EVs incorporate and transport cargoes derived from their parent cells that are relevant for research purposes, as well as for use as diagnostic or prognostic markers in various disease [4]. These include RNA and proteins; however, the average low lumen size of EVs limits the amount of material that they can carry [5,6].…”

Section: Introductionmentioning

confidence: 99%

Proteome profiling of extracellular vesicles captured with the affinity peptide Vn96: comparison of Laemmli and TRIzol© protein‐extraction methods

Joy

Ayre

Chute

et al. 2018

J of Extracellular Vesicle

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Sample amount is often a limiting factor for multi-parametric analyses that encompass at least three areas of ‘-omics’ research: genomics, transcriptomics and proteomics. Limited sample amounts are also an important consideration when these multi-parametric analyses are performed on extracellular vesicles (EVs), as the amount of EVs (and EV cargo) that can be isolated is often very low. It is well understood that a monophasic solution of phenol and guanidine isothiocyanate (i.e. TRIzol©) can simultaneously isolate DNA, RNA and proteins from biological samples; however, it is most commonly used for the extraction of RNA. Validation of this reagent for the isolation of multiple classes of biological molecules from EVs would provide a widely applicable method for performing multi-parametric analyses of EV material. In this report, we describe a comparison of proteins identified from EVs processed with either TRIzol© or the conventional Laemmli buffer protein-extraction reagents. EVs were isolated from 3 mL of cell-culture supernatant derived from MCF-10A, MCF-7 and MDA-MB-231 cells using the Vn96 EV capture technology. For the TRIzol© extraction protocol, proteins were precipitated with acetone from the organic phase and then re-solubilized in a mixture of 8M urea, 0.2% SDS and 1 M Tris-HCl pH 6.8, followed by dilution in 5× loading buffer prior to fractionation with 1D SDS-PAGE. NanoLC-MS/MS of the trypsin-digested proteins was used to generate proteomic profiles from EV protein samples extracted with each method. Of the identified proteins, 57.7%, 69.2% and 57.0% were common to both extraction methods for EVs from MCF-10A, MCF-7 and MDA-MB-231, respectively. Our results suggest that TRIzol© extraction of proteins from EVs has significant equivalence to the traditional Laemmli method. The advantage of using TRIzol© reagent is the ability to accumulate multi-parametric data (e.g., RNA and protein profiles) on the same limited EV sample while minimizing sample preparation and processing time.

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“…They may be protected by extracellular vesicles (19,25). However, the distribution of long RNA in these vesicles is unknown.…”

Section: Introductionmentioning

confidence: 99%

Circulating Long RNAs in Serum Extracellular Vesicles: Their Characterization and Potential Application as Biomarkers for Diagnosis of Colorectal Cancer

Dong

Lin

Pan

et al. 2016

Cancer Epidemiology, Biomarkers &Amp; Prevention

188

111

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Background: Long noncoding RNA (lncRNA) and mRNAs are long RNAs (!200 nucleotides) compared with miRNAs. In blood, long RNAs may be protected by serum extracellular vesicles, such as apoptotic bodies (AB), microvesicles (MV), and exosomes (EXO). They are potential biomarkers for identifying cancer.Methods: Sera from 76 preoperative colorectal cancer patients, 76 age-and sex-matched healthy subjects, and 20 colorectal adenoma patients without colorectal cancer were collected. We investigated the distribution of long RNAs into the three vesicles. Seventy-nine cancer-related long RNAs were chosen and detected using qPCR.Results: The quantity of long RNA has varying distribution among three subtypes of extracellular vesicles in serum. Most mRNA and lncRNA genes had higher quantity in EXOs than that

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Diagnostic and Prognostic Potential of Extracellular Vesicles in Peripheral Blood

Abstract: Overall, the use of EV analysis as a diagnostic and prognostic tool has prodigious clinical potential.

Cited by 225 publications

References 122 publications

Regulation of chronic inflammatory and immune processes by extracellular vesicles

Regulation of chronic inflammatory and immune processes by extracellular vesicles

Proteome profiling of extracellular vesicles captured with the affinity peptide Vn96: comparison of Laemmli and TRIzol© protein‐extraction methods

Circulating Long RNAs in Serum Extracellular Vesicles: Their Characterization and Potential Application as Biomarkers for Diagnosis of Colorectal Cancer

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