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Urine proteomics for discovery of improved diagnostic markers of Kawasaki disease.

Kentsis A, Shulman A, Ahmed S, Brennan E, Monuteaux MC, Lee YH, Lipsett S, Paulo JA, Dedeoglu F, Fuhlbrigge R, Bachur R, Bradwin G, Arditi M, Sundel RP, Newburger JW, Steen H, Kim S - EMBO Mol Med (2012)

Bottom Line: We discovered that urine proteomes of patients with KD, but not those with mimicking conditions, were enriched for markers of cellular injury such as filamin and talin, immune regulators such as complement regulator CSMD3, immune pattern recognition receptor muclin, and immune cytokine protease meprin A.Significant elevations of filamin C and meprin A were detected in both the serum and urine in two independent cohorts of patients with KD, comprised of a total of 236 patients.Notably, meprin A was enriched in the coronary artery lesions of a mouse model of KD.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.

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Patients with Kawasaki disease exhibit a unique urine proteome that is distinct from patients without KD or commonly present urinary proteinsHeatmap of the 15 individual urinary proteomes (columns) showing the results of Bayesian analysis of the top 10 proteins (rows) that are detected in patients with KD as compared to those without. The top 5 proteins that are detected in patients without KD as compared to those with KD are shown for comparison. Blue-to-red colour gradient represents the number of MS/MS spectra (spectral count) that corresponds to urinary protein abundance.Western immunoblot analysis of meprin A and filamin C in urine, demonstrating enrichment of meprin A and filamin C in the urine of patients with KD as compared to patients without KD. Note that meprin A and filamin C are detected predominantly as partial-length isoforms, with apparent molecular weights of 35 and 80 kDa, respectively. Albumin serves as the loading control.
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fig01: Patients with Kawasaki disease exhibit a unique urine proteome that is distinct from patients without KD or commonly present urinary proteinsHeatmap of the 15 individual urinary proteomes (columns) showing the results of Bayesian analysis of the top 10 proteins (rows) that are detected in patients with KD as compared to those without. The top 5 proteins that are detected in patients without KD as compared to those with KD are shown for comparison. Blue-to-red colour gradient represents the number of MS/MS spectra (spectral count) that corresponds to urinary protein abundance.Western immunoblot analysis of meprin A and filamin C in urine, demonstrating enrichment of meprin A and filamin C in the urine of patients with KD as compared to patients without KD. Note that meprin A and filamin C are detected predominantly as partial-length isoforms, with apparent molecular weights of 35 and 80 kDa, respectively. Albumin serves as the loading control.

Mentions: Candidate diagnostic markers of KD were identified based on the analysis of 15 specimens collected at the onset of the study and chosen based on availability: 6 KD specimens (3 without and 3 with coronary artery dilatation), 6 non-KD specimens (2 with non-specific viral syndromes, 3 with adenovirus, and 1 with pyelonephritis), and 3 matched specimens collected from patients with KD 1 month following complete response to treatment (convalescent KD). This analysis identified 2131 unique proteins, with the tissue and physical origin of the aggregate urine proteomes similar to previous studies (Kentsis et al, 2009). Analysis of the three comparison groups led to the identification of more than 190 proteins in the urine of patients with KD, but not in any of the patients without KD or in those whose KD had resolved completely (Supporting Information Table 1). We analysed the abundance of candidate KD markers to identify those that are most enriched in patients with KD, ranking them in order of relative abundance and prevalence (Fig 1). The identified markers include a variety of proteins associated with endothelial and myocardial cell injury such as filamin and titin, and immune regulators such as DMBT1 and meprin A. Many of the detected markers are high molecular weight proteins, which are probably enriched in the urine of patients with KD as processed and/or truncated proteins, as confirmed for meprin A and filamin C by Western immunoblotting (Fig 1A). The discovered proteomes are provided in Supporting Information Table 2 (processed data), and the raw data, in accordance with the MIAPE standard, and are openly available at Peptide Atlas (http://www.peptideatlas.org).


Urine proteomics for discovery of improved diagnostic markers of Kawasaki disease.

Kentsis A, Shulman A, Ahmed S, Brennan E, Monuteaux MC, Lee YH, Lipsett S, Paulo JA, Dedeoglu F, Fuhlbrigge R, Bachur R, Bradwin G, Arditi M, Sundel RP, Newburger JW, Steen H, Kim S - EMBO Mol Med (2012)

Patients with Kawasaki disease exhibit a unique urine proteome that is distinct from patients without KD or commonly present urinary proteinsHeatmap of the 15 individual urinary proteomes (columns) showing the results of Bayesian analysis of the top 10 proteins (rows) that are detected in patients with KD as compared to those without. The top 5 proteins that are detected in patients without KD as compared to those with KD are shown for comparison. Blue-to-red colour gradient represents the number of MS/MS spectra (spectral count) that corresponds to urinary protein abundance.Western immunoblot analysis of meprin A and filamin C in urine, demonstrating enrichment of meprin A and filamin C in the urine of patients with KD as compared to patients without KD. Note that meprin A and filamin C are detected predominantly as partial-length isoforms, with apparent molecular weights of 35 and 80 kDa, respectively. Albumin serves as the loading control.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3569638&req=5

fig01: Patients with Kawasaki disease exhibit a unique urine proteome that is distinct from patients without KD or commonly present urinary proteinsHeatmap of the 15 individual urinary proteomes (columns) showing the results of Bayesian analysis of the top 10 proteins (rows) that are detected in patients with KD as compared to those without. The top 5 proteins that are detected in patients without KD as compared to those with KD are shown for comparison. Blue-to-red colour gradient represents the number of MS/MS spectra (spectral count) that corresponds to urinary protein abundance.Western immunoblot analysis of meprin A and filamin C in urine, demonstrating enrichment of meprin A and filamin C in the urine of patients with KD as compared to patients without KD. Note that meprin A and filamin C are detected predominantly as partial-length isoforms, with apparent molecular weights of 35 and 80 kDa, respectively. Albumin serves as the loading control.
Mentions: Candidate diagnostic markers of KD were identified based on the analysis of 15 specimens collected at the onset of the study and chosen based on availability: 6 KD specimens (3 without and 3 with coronary artery dilatation), 6 non-KD specimens (2 with non-specific viral syndromes, 3 with adenovirus, and 1 with pyelonephritis), and 3 matched specimens collected from patients with KD 1 month following complete response to treatment (convalescent KD). This analysis identified 2131 unique proteins, with the tissue and physical origin of the aggregate urine proteomes similar to previous studies (Kentsis et al, 2009). Analysis of the three comparison groups led to the identification of more than 190 proteins in the urine of patients with KD, but not in any of the patients without KD or in those whose KD had resolved completely (Supporting Information Table 1). We analysed the abundance of candidate KD markers to identify those that are most enriched in patients with KD, ranking them in order of relative abundance and prevalence (Fig 1). The identified markers include a variety of proteins associated with endothelial and myocardial cell injury such as filamin and titin, and immune regulators such as DMBT1 and meprin A. Many of the detected markers are high molecular weight proteins, which are probably enriched in the urine of patients with KD as processed and/or truncated proteins, as confirmed for meprin A and filamin C by Western immunoblotting (Fig 1A). The discovered proteomes are provided in Supporting Information Table 2 (processed data), and the raw data, in accordance with the MIAPE standard, and are openly available at Peptide Atlas (http://www.peptideatlas.org).

Bottom Line: We discovered that urine proteomes of patients with KD, but not those with mimicking conditions, were enriched for markers of cellular injury such as filamin and talin, immune regulators such as complement regulator CSMD3, immune pattern recognition receptor muclin, and immune cytokine protease meprin A.Significant elevations of filamin C and meprin A were detected in both the serum and urine in two independent cohorts of patients with KD, comprised of a total of 236 patients.Notably, meprin A was enriched in the coronary artery lesions of a mouse model of KD.

View Article: PubMed Central - PubMed

Affiliation: Division of Hematology/Oncology, Department of Pediatric Oncology, Dana-Farber Cancer Institute, Boston Children's Hospital, Harvard Medical School, Boston, MA, USA.

Show MeSH
Related in: MedlinePlus