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  • Review Article
  • Published:

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

Nature Reviews Nephrology volume 13, pages 731–749 (2017)Cite this article

Subjects

Key Points

  • Urinary extracellular vesicles comprise a wide range of biologically distinct structures with contents that are a snapshot of the life of a cell

  • Urine is a dynamic biofluid, which changes over hours and days within an individual; therefore, at present, no single approach for the isolation of urinary extracellular vesicles is likely to comprehensively distinguish between healthy and disease states

  • Alterations in the composition of urinary extracellular vesicles are useful experimentally and may provide information about disease pathophysiology as well as provide diagnostic end points for the study of renal disease

  • Perhaps the greatest promise of this 'extracellular organelle' is to open a window for science into a greater understanding of cellular therapeutics

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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Figure 1: Mechanisms of urinary extracellular vesicle formation regulate their composition.
Figure 2: Overview of exosome formation.
Figure 3: Comparison of approaches to isolate extracellular vesicles from the urine of healthy individuals and from patients with nephrotic syndrome.

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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).

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Authors and Affiliations

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

    Michael L. Merchant & Jon B. Klein

  2. Department of Nephrology, Radboud University Medical Center, Radboud Institute of Health Science, Nijmegen, 6500HB, The Netherlands

    Ilse M. Rood & Jeroen K. J. Deegens

  3. Robley Rex Veterans Administration Medical Center, Louisville, 40206, Kentucky, USA

    Jon B. Klein

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  1. Michael L. Merchant
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  2. Ilse M. Rood
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  3. Jeroen K. J. Deegens
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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.

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Correspondence to Jon B. Klein.

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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.

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Merchant, M., Rood, I., Deegens, J. et al. Isolation and characterization of urinary extracellular vesicles: implications for biomarker discovery. Nat Rev Nephrol 13, 731–749 (2017). https://doi.org/10.1038/nrneph.2017.148

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