Comparative Study of Extracellular Vesicles from the Urine of Healthy Individuals and Prostate Cancer Patients

Recent studies suggest that extracellular vesicles may be the key to timely diagnosis and monitoring of genito-urological malignancies. In this study we investigated the composition and content of extracellular vesicles found in the urine of healthy donors and prostate cancer patients. Urine of 14 P...

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Published in:	PloS one Vol. 11; no. 6; p. e0157566
Main Authors:	Bryzgunova, Olga E, Zaripov, Marat M, Skvortsova, Tatyana E, Lekchnov, Evgeny A, Grigor'eva, Alina E, Zaporozhchenko, Ivan A, Morozkin, Evgeny S, Ryabchikova, Elena I, Yurchenko, Yuri B, Voitsitskiy, Vladimir E, Laktionov, Pavel P
Format:	Journal Article
Language:	English
Published:	United States Public Library of Science 15-06-2016 Public Library of Science (PLoS)
Subjects:	Aged Aged, 80 and over Biology Biology and Life Sciences Biomarkers Cancer Centrifugation Centrifugation - methods Clinics Comparative studies Deoxyribonucleic acid Diagnostic systems DNA DNA, Neoplasm - genetics DNA, Neoplasm - urine Electron microscopy Enrichment Exosomes Exosomes - metabolism Exosomes - ultrastructure Extracellular vesicles Extracellular Vesicles - metabolism Extracellular Vesicles - ultrastructure Humans Laboratories Male Medical diagnosis Medicine Medicine and Health Sciences MicroRNAs MicroRNAs - genetics MicroRNAs - urine Microscopy Microscopy, Electron, Transmission Middle Aged Mitochondrial DNA Neoplasm Proteins - metabolism Neoplasm Proteins - urine Oncology Patients Prostate cancer Prostatic Neoplasms - diagnosis Prostatic Neoplasms - urine Proteins Proteomics Radiation therapy Research and Analysis Methods Reverse Transcriptase Polymerase Chain Reaction Ribonucleic acid RNA RNA, Neoplasm - genetics RNA, Neoplasm - urine Sensitivity and Specificity Signal transduction Size distribution Transmission electron microscopy Ultrafiltration Urine Vesicles
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Summary:	Recent studies suggest that extracellular vesicles may be the key to timely diagnosis and monitoring of genito-urological malignancies. In this study we investigated the composition and content of extracellular vesicles found in the urine of healthy donors and prostate cancer patients. Urine of 14 PCa patients and 20 healthy volunteers was clarified by low-speed centrifugation and total extracellular vesicles fraction was obtain by high-speed centrifugation. The exosome-enriched fraction was obtained by filtration of total extracellular vesicles through a 0.1 μm pore filter. Transmission electron microscopy showed that cell-free urine in both groups contained vesicles from 20 to 230 nm. Immunogold staining after ultrafiltration demonstrated that 95% and 90% of extracellular vesicles in healthy individuals and cancer patients, respectively, were exosomes. Protein, DNA and RNA concentrations as well as size distribution of extracellular vesicles in both fractions were analyzed. Only 75% of the total protein content of extracellular vesicles was associated with exosomes which amounted to 90-95% of all vesicles. Median DNA concentrations in total extracellular vesicles and exosome-enriched fractions were 18 pg/ml and 2.6 pg/ml urine, correspondingly. Urine extracellular vesicles carried a population of RNA molecules 25 nt to 200 nt in concentration of no more than 290 pg/ml of urine. Additionally, concentrations of miR-19b, miR-25, miR-125b, and miR-205 were quantified by qRT-PCR. MiRNAs were shown to be differently distributed between different fractions of extracellular vesicles. Detection of miR-19b versus miR-16 in total vesicles and exosome-enriched fractions achieved 100%/93% and 95%/79% specificity/sensitivity in distinguishing cancer patients from healthy individuals, respectively, demonstrating the diagnostic value of urine extracellular vesicles.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Competing Interests: The authors have declared that no competing interests exist. Conceived and designed the experiments: OEB IAZ ESM PPL. Performed the experiments: OEB MMZ TES EAL IAZ AEG EIR. Analyzed the data: OEB IAZ PPL. Contributed reagents/materials/analysis tools: AEG EIR. Wrote the paper: OEB EAL IAZ ESM PPL. Acqusition of clinical samples: VEV MMZ YBY.
ISSN:	1932-6203 1932-6203
DOI:	10.1371/journal.pone.0157566