Review

Isolation and characterization of urinary extracellular vesicles: implications for biomarker discovery

  • Nature Reviews Nephrology 13, 731749 (2017)
  • doi:10.1038/nrneph.2017.148
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Abstract

Urine is a valuable diagnostic medium and, with the discovery of urinary extracellular vesicles, is viewed as a dynamic bioactive fluid. Extracellular vesicles are lipid-enclosed structures that can be classified into three categories: exosomes, microvesicles (or ectosomes) and apoptotic bodies. This classification is based on the mechanisms by which membrane vesicles are formed: fusion of multivesicular bodies with the plasma membranes (exosomes), budding of vesicles directly from the plasma membrane (microvesicles) or those shed from dying cells (apoptotic bodies). During their formation, urinary extracellular vesicles incorporate various cell-specific components (proteins, lipids and nucleic acids) that can be transferred to target cells. The rigour needed for comparative studies has fueled the search for optimal approaches for their isolation, purification, and characterization. RNA, the newest extracellular vesicle component to be discovered, has received substantial attention as an extracellular vesicle therapeutic, and compelling evidence suggests that ex vivo manipulation of microRNA composition may have uses in the treatment of kidney disorders. The results of these studies are building the case that urinary extracellular vesicles act as mediators of renal pathophysiology. As the field of extracellular vesicle studies is burgeoning, this Review focuses on primary data obtained from studies of human urine rather than on data from studies of laboratory animals or cultured immortalized cells.

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Acknowledgements

J.B.K. and M.L.M. are supported in part by grants R01DK110077 (J.B.K.), R01DK091584 (M.L.M.), P50AA024337 (M.L.M.) and P20GM113226 (MLM) from the US NIH. I.M.R. is supported by an AGIKO grant from ZonMw-NWO (grant 92003587).

Author information

Affiliations

  1. Division of Nephrology and Hypertension, Department of Medicine, University of Louisville, Louisville, Kentucky 40202, USA.

    • Michael L. Merchant
    •  & Jon B. Klein
  2. Radboud University Medical Center, Radboud Institute of Health Science, Department of Nephrology, Nijmegen 6500HB, The Netherlands.

    • Ilse M. Rood
    •  & Jeroen K. J. Deegens
  3. Robley Rex Veterans Administration Medical Center, Louisville, Kentucky 40206, USA.

    • Jon B. Klein

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Contributions

All authors researched data for the article. M.L.M, I.M.R, J.K.J.D. and J.B.K made substantial contributions to discussions of the content. M.L.M, I.M.R and J.K.J.D. wrote the article and M.L.M., I.M.R., J.K.J.D. and J.B.K. reviewed and/or edited the manuscript before submission.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Jon B. Klein.

Glossary

mRNAs

Messenger RNAs, which are transcripts of DNA.

MicroRNAs

(miRNAs). Small, non-coding RNAs that regulate gene expression post-transcriptionally by targeting specific mRNAs for inhibition or degradation through complementary base pairing.

Exosomes

Extracellular vesicles that are formed by inward budding of the cell membrane, followed by fusion with a multivesicular body (MVB) and formation of intraluminal vesicles inside the MVB. The intraluminal vesicles that are released by fusion of the MVB with the cell plasma membrane are called exosomes.

Microvesicles

Extracellular vesicles that are formed by direct budding from the cell plasma membrane.

Apoptotic bodies

Extracellular vesicle that are released during the late stages of cell death.

Transcriptomics

The study of the complete set of RNA transcripts (the transcriptome) that is encoded by the genome, underspecific circumstances or in a specific cell.

Proteomics

Large scale studies of proteins involving the systematic identification and quantification of the complete set of proteins (the proteome) of a biological system (cell, tissue, organ, biological fluid or organism) at a specific point in time. Mass spectrometry is the technique most often used for proteomic analysis.

Nephrotic syndrome

A constellation of symptoms characterized by heavy proteinuria (>3–3.5 g/24 h), hypoalbuminuria (<30 g/l), peripheral oedema and hyperlipidaemia.