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iTRAQ proteomic analysis of extracellular matrix remodeling in aortic valve disease.

Martin-Rojas T, Mourino-Alvarez L, Alonso-Orgaz S, Rosello-Lleti E, Calvo E, Lopez-Almodovar LF, Rivera M, Padial LR, Lopez JA, de la Cuesta F, Barderas MG - Sci Rep (2015)

Bottom Line: Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms.The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses.These findings are substantiated by our previous results demonstrating differential ECM protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Physiopathology, Hospital Nacional de Parapléjicos, SESCAM, Toledo, Spain.

ABSTRACT
Degenerative aortic stenosis (AS) is the most common worldwide cause of valve replacement. The aortic valve is a thin, complex, layered connective tissue with compartmentalized extracellular matrix (ECM) produced by specialized cell types, which directs blood flow in one direction through the heart. There is evidence suggesting remodeling of such ECM during aortic stenosis development. Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms. Aortic valve samples were collected from 18 patients which underwent aortic valve replacement (50% males, mean age of 74 years) and 18 normal control valves were obtained from necropsies (40% males, mean age of 69 years). The proteome of the samples was analyzed by 2D-LC MS/MS iTRAQ methodology. The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses. These findings are substantiated by our previous results demonstrating differential ECM protein expression. The present study has demonstrated a differential ECM protein pattern in individuals with AS, therefore supporting previous evidence of a dynamic ECM remodeling in human aortic valves during AS development.

No MeSH data available.


Related in: MedlinePlus

Validation of differential proteins by SRM analysis.Graphical representation of the abundance (left panels) and representative extracted ion chromatograms (right panels) of the transitions observed for periostin (A) and prolargin (B). Significant increases (p < 0.05) in the expression of the measured proteotypic peptides of both proteins were observed in AS with respect to healthy valves. *p < 0.05; **p < 0.01. C: controls; AS: aortic stenosis; cps: counts per second.
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f4: Validation of differential proteins by SRM analysis.Graphical representation of the abundance (left panels) and representative extracted ion chromatograms (right panels) of the transitions observed for periostin (A) and prolargin (B). Significant increases (p < 0.05) in the expression of the measured proteotypic peptides of both proteins were observed in AS with respect to healthy valves. *p < 0.05; **p < 0.01. C: controls; AS: aortic stenosis; cps: counts per second.

Mentions: For each protein, two peptides were quantified by SRM; in the case of periostin and prolargin, these peptides were found to differ significantly in their expression between groups (Fig. 4; Panel A = periostin, peptide 1: p = 0.0022 and peptide 2: p = 0.0053 and panel B = prolargin, peptide1: p = 0.0086 and peptide 2: p = 0.0481) Unfortunately, we were not able to detect the peptides that correspond to biglycan by this technique.


iTRAQ proteomic analysis of extracellular matrix remodeling in aortic valve disease.

Martin-Rojas T, Mourino-Alvarez L, Alonso-Orgaz S, Rosello-Lleti E, Calvo E, Lopez-Almodovar LF, Rivera M, Padial LR, Lopez JA, de la Cuesta F, Barderas MG - Sci Rep (2015)

Validation of differential proteins by SRM analysis.Graphical representation of the abundance (left panels) and representative extracted ion chromatograms (right panels) of the transitions observed for periostin (A) and prolargin (B). Significant increases (p < 0.05) in the expression of the measured proteotypic peptides of both proteins were observed in AS with respect to healthy valves. *p < 0.05; **p < 0.01. C: controls; AS: aortic stenosis; cps: counts per second.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4664895&req=5

f4: Validation of differential proteins by SRM analysis.Graphical representation of the abundance (left panels) and representative extracted ion chromatograms (right panels) of the transitions observed for periostin (A) and prolargin (B). Significant increases (p < 0.05) in the expression of the measured proteotypic peptides of both proteins were observed in AS with respect to healthy valves. *p < 0.05; **p < 0.01. C: controls; AS: aortic stenosis; cps: counts per second.
Mentions: For each protein, two peptides were quantified by SRM; in the case of periostin and prolargin, these peptides were found to differ significantly in their expression between groups (Fig. 4; Panel A = periostin, peptide 1: p = 0.0022 and peptide 2: p = 0.0053 and panel B = prolargin, peptide1: p = 0.0086 and peptide 2: p = 0.0481) Unfortunately, we were not able to detect the peptides that correspond to biglycan by this technique.

Bottom Line: Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms.The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses.These findings are substantiated by our previous results demonstrating differential ECM protein expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Vascular Physiopathology, Hospital Nacional de Parapléjicos, SESCAM, Toledo, Spain.

ABSTRACT
Degenerative aortic stenosis (AS) is the most common worldwide cause of valve replacement. The aortic valve is a thin, complex, layered connective tissue with compartmentalized extracellular matrix (ECM) produced by specialized cell types, which directs blood flow in one direction through the heart. There is evidence suggesting remodeling of such ECM during aortic stenosis development. Thus, a better characterization of the role of ECM proteins in this disease would increase our understanding of the underlying molecular mechanisms. Aortic valve samples were collected from 18 patients which underwent aortic valve replacement (50% males, mean age of 74 years) and 18 normal control valves were obtained from necropsies (40% males, mean age of 69 years). The proteome of the samples was analyzed by 2D-LC MS/MS iTRAQ methodology. The results showed an altered expression of 13 ECM proteins of which 3 (biglycan, periostin, prolargin) were validated by Western blotting and/or SRM analyses. These findings are substantiated by our previous results demonstrating differential ECM protein expression. The present study has demonstrated a differential ECM protein pattern in individuals with AS, therefore supporting previous evidence of a dynamic ECM remodeling in human aortic valves during AS development.

No MeSH data available.


Related in: MedlinePlus