Osteoprotegerin in Exosome-Like Vesicles from Human Cultured Tubular Cells and Urine
Entity
UAM. Departamento de MedicinaPublisher
Public Library of ScienceDate
2013-08-23Citation
10.1371/journal.pone.0072387
PLoS One 8.8 (2013): e72387
ISSN
1932-6203DOI
10.1371/journal.pone.0072387Funded by
This work was supported by grants from the Instituto de Salud Carlos III (ISCIIIRETIC REDINREN RD06/0016, RD12/0021, PI11/01854, PI10/00072 PI09/ 00641 and PS09/00447); Comunidad de Madrid (Fibroteam S2010/BMD-2321, S2010/BMD-2378); Sociedad Española de NefrologÍa; European Network (HEALTH F2-2008-200647); DIALOK European project LSHB-CT-2007-036644; Fundacion Lilly and IRSIN/FRIAT to JE; Programa Intensificación Actividad Investigadora (ISCIII/ Agencia Laín-Entralgo/CM) to AO; Instituto de Salud Carlos III (FIS PI11/01401, CP09/00229); and Fundación Conchita Rábago de Jiménez DÍaz to GAL. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscriptProject
Comunidad de Madrid. S2010/BMD-2378/CIFRA; Comunidad de Madrid. S2010/BMD-2321/FIBROTEAMEditor's Version
http://dx.doi.org/10.1371/journal.pone.0072387Subjects
Apoptosis; Cell membranes; Epithelial cells; Kidneys; Menbrane proteins; Urine; vesicles; Extracellular matrix proteins; MedicinaAbstract
Urinary exosomes have been proposed as potential diagnostic tools. TNF superfamily cytokines and receptors may be
present in exosomes and are expressed by proximal tubular cells. We have now studied the expression of selected TNF
superfamily proteins in exosome-like vesicles from cultured human proximal tubular cells and human urine and have
identified additional proteins in these vesicles by LC-MS/MS proteomics. Human proximal tubular cells constitutively
released exosome-like vesicles that did not contain the TNF superfamily cytokines TRAIL or TWEAK. However, exosome-like
vesicles contained osteoprotegerin (OPG), a TNF receptor superfamily protein, as assessed by Western blot, ELISA or
selected reaction monitoring by nLC-(QQQ)MS/MS. Twenty-one additional proteins were identified in tubular cell exosomelike
vesicles, including one (vitamin D binding protein) that had not been previously reported in exosome-like vesicles.
Twelve were extracellular matrix proteins, including the basement membrane proteins type IV collagen, nidogen-1, agrin
and fibulin-1. Urine from chronic kidney disease patients contained a higher amount of exosomal protein and exosomal
OPG than urine from healthy volunteers. Specifically OPG was increased in autosomal dominant polycystic kidney disease
urinary exosome-like vesicles and expressed by cystic epithelium in vivo. In conclusion, OPG is present in exosome-like
vesicles secreted by proximal tubular epithelial cells and isolated from Chronic Kidney Disease urine.
Files in this item
Google Scholar:Benito-Martin, Alberto
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Conrado Ucero, Álvaro
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Zubiri, Irene
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Posada-Ayala, María
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Fernández-Fernández, Beatriz
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Cannata, Pablo
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Sánchez Niño, María Dolores
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Ruiz Ortega, Marta
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Egido, Jesús
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Álvarez-Llamas, Gloria
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Ortiz Arduán, Alberto
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- Producción científica de la UAM [20684]
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