Secreted proteins from carotid endarterectomy: an untargeted approach to disclose molecular clues of plaque progression

Journal of Translational Medicine

Antonella Cecchettini¹  Maria Giovanna Trivella²  Mauro Ferrari³  Lorenzo Citti²  Federica Viglione²  Michele Marconi³  Silvia Rosini²  Gualtiero Pelosi²  Silvia Rocchiccioli² 

[1] University of Pisa, Department of Clinical and Experimental Medicine, Via Volta, Pisa, Italy;National Research Council, Institute of Clinical Physiology, Via Moruzzi, Pisa, Italy;University of Pisa, Unit of Vascular Surgery, Via Paradisa, Pisa, Italy

关键词: Secretome;    Proteomics;    Carotid plaques;    Atherosclerosis;   

Others  :  825055
DOI  :  10.1186/1479-5876-11-260

 received in 2013-07-10, accepted in 2013-09-18,  发布年份 2013

【 摘 要 】

Background

Atherosclerosis is the main cause of morbidity and mortality in Western countries and carotid plaque rupture is associated to acute events and responsible of 15-20% of all ischemic strokes. Several proteomics approaches have been up to now used to elucidate the molecular mechanisms involved in plaque formation as well as to identify markers of pathology severity for early diagnosis or target of therapy. The aim of this study was to characterize the plaque secretome. The advantage of this approach is that secretome mimics the in vivo condition and implies a reduced complexity compared to the whole tissue proteomics allowing the detection of under-represented potential biomarkers.

Methods

Secretomes from carotid endarterectomy specimens of 14 patients were analyzed by a liquid chromatography approach coupled with label free mass spectrometry. Differential expression of proteins released from plaques and from their downstream distal side segments were evaluated in each specimen. Results were validated by Western blot analysis and ELISA assays. Histology and immunohistochemistry were performed to characterize plaques and to localise the molecular factors highlighted by proteomics.

Results

A total of 463 proteins were identified and 31 proteins resulted differentially secreted from plaques and corresponding downstream segments. A clear-cut distinction in the distribution of cellular- and extracellular-derived proteins, evidently related to the higher cellularity of distal side segments, was observed along the longitudinal axis of carotid endarterectomy samples. The expressions of thrombospondin-1, vitamin D binding protein, and vinculin, as examples of extracellular and intracellular proteins, were immunohistologically compared between adjacent segments and validated by antibody assays. ELISA assays of plasma samples from 34 patients and 10 healthy volunteers confirmed a significantly higher concentration of thrombospondin-1 and vitamin D binding protein in atherosclerotic subjects.

Conclusions

Taking advantage of the optimized workflow, a detailed protein profile related to carotid plaque secretome has been produced which may assist and improve biomarker discovery of molecular factors in blood. Distinctive signatures of proteins secreted by adjacent segments of carotid plaques were evidenced and they may help discriminating markers of plaque complication from those of plaque growth.

【 授权许可】

   
2013 Rocchiccioli et al.; licensee BioMed Central Ltd.

【 图 表 】

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