Differential protein profiling of renal cell carcinoma urinary exosomes

F. Raimondo, L. Morosi, S. Corbetta, C. Chinello, P. Brambilla, P. Della Mina, A. Villa, G. Albo, C. Battaglia, S. Bosari, F. Magni, M. Pitto

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

Renal cell carcinoma (RCC) accounts for about 3% of all human malignancies and its incidence is increasing. There are no standard biomarkers currently used in the clinical management of patients with renal cell carcinoma. A promising strategy for new biomarker detection is comparative proteomics of urinary exosomes (UE), nanovesicles released by every epithelial cell facing the urinary space, enriched in renal proteins and excluding high-abundance plasmatic proteins, such as albumin. Aim of the work is to establish the protein profile of exosomes isolated from urines of RCC patient compared with control subjects. We enrolled 29 clear cell RCC patients and 23 control healthy subjects (CTRL), age and sex-matched, for urine collection and vesicle isolation by differential centrifugation. Such vesicles were morphologically and biochemically characterized and proved to share exosome properties. Proteomic analysis, performed on 9 urinary exosome (UE) pooled samples by gel based digestion followed by LC-MS/MS, led to the identification of 261 proteins from CTRL subject UE and 186 from RCC patient UE, and demonstrated that most of the identified proteins are membrane associated or cytoplasmic. Moreover, about a half of identified proteins are not shared between RCC and control UE. Starting from these observations, and from the literature, we selected a panel of 10 proteins, whose UE differential content was subjected to immunoblotting validation. Results show for the first time that RCC UE protein content is substantially and reproducibly different from control UE, and that these differences may provide clues for new RCC biomarker discovery.

Original languageEnglish
Pages (from-to)1220-1233
Number of pages14
JournalMolecular BioSystems
Volume9
Issue number6
DOIs
Publication statusPublished - 2013

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Exosomes
Renal Cell Carcinoma
Proteins
Biomarkers
Proteomics
Urine Specimen Collection
Centrifugation
Immunoblotting
Digestion
Albumins
Healthy Volunteers
Membrane Proteins
Gels
Epithelial Cells
Urine

ASJC Scopus subject areas

  • Biotechnology
  • Molecular Biology
  • Medicine(all)

Cite this

Raimondo, F., Morosi, L., Corbetta, S., Chinello, C., Brambilla, P., Della Mina, P., ... Pitto, M. (2013). Differential protein profiling of renal cell carcinoma urinary exosomes. Molecular BioSystems, 9(6), 1220-1233. https://doi.org/10.1039/c3mb25582d

Differential protein profiling of renal cell carcinoma urinary exosomes. / Raimondo, F.; Morosi, L.; Corbetta, S.; Chinello, C.; Brambilla, P.; Della Mina, P.; Villa, A.; Albo, G.; Battaglia, C.; Bosari, S.; Magni, F.; Pitto, M.

In: Molecular BioSystems, Vol. 9, No. 6, 2013, p. 1220-1233.

Research output: Contribution to journalArticle

Raimondo, F, Morosi, L, Corbetta, S, Chinello, C, Brambilla, P, Della Mina, P, Villa, A, Albo, G, Battaglia, C, Bosari, S, Magni, F & Pitto, M 2013, 'Differential protein profiling of renal cell carcinoma urinary exosomes', Molecular BioSystems, vol. 9, no. 6, pp. 1220-1233. https://doi.org/10.1039/c3mb25582d
Raimondo F, Morosi L, Corbetta S, Chinello C, Brambilla P, Della Mina P et al. Differential protein profiling of renal cell carcinoma urinary exosomes. Molecular BioSystems. 2013;9(6):1220-1233. https://doi.org/10.1039/c3mb25582d
Raimondo, F. ; Morosi, L. ; Corbetta, S. ; Chinello, C. ; Brambilla, P. ; Della Mina, P. ; Villa, A. ; Albo, G. ; Battaglia, C. ; Bosari, S. ; Magni, F. ; Pitto, M. / Differential protein profiling of renal cell carcinoma urinary exosomes. In: Molecular BioSystems. 2013 ; Vol. 9, No. 6. pp. 1220-1233.
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