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A pipeline that integrates the discovery and verification of plasma protein biomarkers reveals candidate markers for cardiovascular disease
TA Addona, X Shi, H Keshishian, DR Mani... - Nature ..., 2011 - We developed a pipeline to integrate the proteomic technologies used from the discovery to
the verification stages of plasma biomarker identification and applied it to identify early biomarkers
of cardiac injury from the blood of patients undergoing a therapeutic, planned myocardial ...
We developed a pipeline to integrate the proteomic technologies used from the discovery to the verification stages of plasma biomarker identification and applied it to identify early biomarkers of cardiac injury from the blood of patients undergoing a therapeutic, planned myocardial infarction (PMI) for treatment of hypertrophic cardiomyopathy. Sampling of blood directly from patient hearts before, during and after controlled myocardial injury ensured enrichment for candidate biomarkers and allowed patients to serve as their own biological controls. LC-MS/MS analyses detected 121 highly differentially expressed proteins, including previously credentialed markers of cardiovascular disease and >100 novel candidate biomarkers for myocardial infarction (MI). Accurate inclusion mass screening (AIMS) qualified a subset of the candidates based on highly specific, targeted detection in peripheral plasma, including some markers unlikely to have been identified without this step. Analyses of peripheral plasma from controls and patients with PMI or spontaneous MI by quantitative multiple reaction monitoring mass spectrometry or immunoassays suggest that the candidate biomarkers may be specific to MI. This study demonstrates that modern proteomic technologies, when coherently integrated, can yield novel cardiovascular biomarkers meriting further evaluation in large, heterogeneous cohorts.
Blood samples were collected from the coronary sinus of patients undergoing alcohol septal ablation for hypertrophic cardiomyopathy (planned myocardial infarction or PMI) before ablation (baseline),and at 10 and 60 min after ablation. The coronary sinus samples, comprising a proximal fluid of the heart, were used for discovery proteomics, using abundant protein depletion and extensive fractionation and LC-MS/MS of peptides to generate a prioritized list of biomarker candidates. Peripheral blood was collected from patients undergoing PMI at the same time points and additional time points extending to 24 h after ablation. Blood collected up to 4 h after ablation was used for analytical qualification by AIMS, a process that determines which of the differentially abundant proteins from the discovery experiments are detectable in peripheral blood. Qualified protein biomarker candidates were subsequently quantitatively measured in peripheral blood using immunoassays (when antibodies were available) and SID-MRM-MS when antibody reagents were not available. Figure was adapted from reference 29. Blue type indicates proximal fluid. Red type indicates peripheral blood.
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A pipeline that integrates the discovery and verification of plasma protein biomarkers reveals candidate markers for cardiovascular disease |
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