Unravelling the proteomic landscape of extracellular vesicles in prostate cancer by density-based fractionation of urine
Dhondt, Bert; Geeurickx, Edward; Tulkens, Joeri; Van Deun, Jan; Vergauwen, Glenn; Lippens, Lien; Miinalainen, Ilkka; Rappu, Pekka; Heino, Jyrki; Ost, Piet; Lumen, Nicolaas; De Wever, Olivier; Hendrix, An (2020-03-11)
Bert Dhondt, Edward Geeurickx, Joeri Tulkens, Jan Van Deun, Glenn Vergauwen, Lien Lippens, Ilkka Miinalainen, Pekka Rappu, Jyrki Heino, Piet Ost, Nicolaas Lumen, Olivier De Wever & An Hendrix (2020) Unravelling the proteomic landscape of extracellular vesicles in prostate cancer by density-based fractionation of urine, Journal of Extracellular Vesicles, 9:1, DOI: 10.1080/20013078.2020.1736935
© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
https://creativecommons.org/licenses/by-nc/4.0/
https://urn.fi/URN:NBN:fi-fe2020062245268
Tiivistelmä
Abstract
Extracellular vesicles (EV) are increasingly being recognized as important vehicles of intercellular communication and promising diagnostic and prognostic biomarkers in cancer. Despite this enormous clinical potential, the plethora of methods to separate EV from biofluids, providing material of highly variable purity, and lacking knowledge regarding methodological repeatability pose a barrier to clinical translation. Urine is considered an ideal proximal fluid for the study of EV in urological cancers due to its direct contact with the urogenital system. We demonstrate that density-based fractionation of urine by bottom-up Optiprep density gradient centrifugation separates EV and soluble proteins with high specificity and repeatability. Mass spectrometry-based proteomic analysis of urinary EV (uEV) in men with benign and malignant prostate disease allowed us to significantly expand the known human uEV proteome with high specificity and identifies a unique biological profile in prostate cancer not uncovered by the analysis of soluble proteins. In addition, profiling the proteome of EV separated from prostate tumour conditioned medium and matched uEV confirms the specificity of the identified uEV proteome for prostate cancer. Finally, a comparative proteomic analysis with uEV from patients with bladder and renal cancer provided additional evidence of the selective enrichment of protein signatures in uEV reflecting their respective cancer tissues of origin. In conclusion, this study identifies hundreds of previously undetected proteins in uEV of prostate cancer patients and provides a powerful toolbox to map uEV content and contaminants ultimately allowing biomarker discovery in urological cancers.
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