Abstract
In the last years, disease biomarker discovery has highly evolved thanks to the application of high-throughput technologies such as proteomics. However, due to the elevated complexity and abundance of some of the proteins in the samples the analysis of subcellular compartments has been revealed to be fundamental in order to identify underrepresented clinically relevant proteins. In this sense, extracellular microvesicles including exosomes that are present in different body fluids constitute a suitable and convenient subcellular compartment to identify disease biomarkers. On the other hand, animal models offer numerous advantages over human samples in order to accelerate the identification of candidate biomarkers. In this chapter we provide a detailed methodology to purify and analyze urinary exosomes that can be applied to the study of different diseases that have good animal models.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Simons M, Raposo G (2009) Exosomes – vesicular carriers for intercellular communication. Curr Opin Cell Biol 21(4):575–581 [Pubmed: 19442504]
Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO (2007) Exosome-mediated transfer of mRNAs and microRNAs is a novel mechanism of genetic exchange between cells. Nat Cell Biol 9(6):654–659 [Pubmed: 17486113]
Nguyen DG, Booth A, Gould SJ, Hildreth JE (2003) Evidence that HIV budding in primary macrophages occurs through the exosome release pathway. J Biol Chem 278(52):52347–52354 [Pubmed: 14561735]
Andre F, Escudier B, Angevin E, Tursz T, Zitvogel L (2004) Exosomes for cancer immunotherapy. Ann Oncol 15(Suppl 4):iv141–iv144 [Pubmed: 15477298]
Mathivanan S, Ji H, Simpson RJ (2010) Exosomes: extracellular organelles important in intercellular communication. J Proteomics 73(10):1907–1920 [Pubmed: 20601276]
Simpson RJ, Lim JW, Moritz RL, Mathivanan S (2009) Exosomes: proteomic insights and diagnostic potential. Expert Rev Proteomics 6(3):267–283 [Pubmed: 19489699]
Pisitkun T, Shen RF, Knepper MA (2004) Identification and proteomic profiling of exosomes in human urine. Proc Natl Acad Sci USA 101(36):13368–13373 [Pubmed: 15326289]
Gonzales PA, Pisitkun T, Hoffert JD et al (2009) Large-scale proteomics and phosphoproteomics of urinary exosomes. J Am Soc Nephrol 20(2):363–379 [Pubmed: 19056867]
Zhou H, Cheruvanky A, Hu X et al (2008) Urinary exosomal transcription factors, a new class of biomarkers for renal disease. Kidney Int 74(5):613–621 [Pubmed: 18509321]
Mitchell PJ, Welton J, Staffurth J et al (2009) Can urinary exosomes act as treatment response markers in prostate cancer? J Transl Med 7:4 [Pubmed: 19138409]
Schughart K (2010) SYSGENET: a meeting report from a new European network for systems genetics. Mamm Genome 21(7–8):331–336 [Pubmed: 20623354]
Conde-Vancells J, Rodriguez-Suarez E, Gonzalez E et al (2010) Candidate biomarkers in exosome-like vesicles purified from rat and mouse urine samples. Proteomics Clin Appl 4(4):416–425 [Pubmed: 20535238]
Keppler D, Lesch R, Reutter W, Decker K (1968) Experimental hepatitis induced by d-galactosamine. Exp Mol Pathol 9(2):279–290 [Pubmed: 4952077]
Lu SC, Alvarez L, Huang ZZ et al (2001) Methionine adenosyltransferase 1A knockout mice are predisposed to liver injury and exhibit increased expression of genes involved in proliferation. Proc Natl Acad Sci USA 98(10):5560–5565 [Pubmed: 11320206]
Martinez-Chantar ML, Vazquez-Chantada M, Ariz U et al (2008) Loss of the glycine N-methyltransferase gene leads to steatosis and hepatocellular carcinoma in mice. Hepatology 47(4):1191–1199 [Pubmed: 18318442]
Cocucci E, Racchetti G, Meldolesi J (2009) Shedding microvesicles: artefacts no more. Trends Cell Biol 19(2):43–51 [Pubmed: 19144520]
Pisitkun T, Johnstone R, Knepper MA (2006) Discovery of urinary biomarkers. Mol Cell Proteomics 5(10):1760–1771 [Pubmed: 16837576]
Acknowledgements
This work was supported by grants from Foundation the Institute of Health Carlos III (06/0621 and 09/00526 to JMFP), the program Ramon y Cajal (JMFP). Centro de Investigación Biomédica en Red en el Área temática de Enfermedades Hepáticas y Digestivas (CIBERehd) is funded by the Institute of Health Carlos III.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2012 Springer Science+Business Media New York
About this protocol
Cite this protocol
Conde-Vancells, J., Falcon-Perez, J.M. (2012). Isolation of Urinary Exosomes from Animal Models to Unravel Noninvasive Disease Biomarkers. In: Josic, D., Hixson, D. (eds) Liver Proteomics. Methods in Molecular Biology, vol 909. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-959-4_21
Download citation
DOI: https://doi.org/10.1007/978-1-61779-959-4_21
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-61779-958-7
Online ISBN: 978-1-61779-959-4
eBook Packages: Springer Protocols