title: Aptamer-Based Proteomic Profiling Reveals Novel Candidate Biomarkers and Pathways in Cardiovascular Disease.
abstract: Single-stranded DNA aptamers are oligonucleotides of ≈50 base pairs in length selected for their ability to bind proteins with high specificity and affinity. Emerging DNA aptamer-based technologies may address limitations of existing proteomic techniques, including low sample throughput, which have hindered proteomic analyses of large cohorts.
  
  
  
  To identify early biomarkers of myocardial injury, we applied an aptamer-based proteomic platform that measures 1129 proteins to a clinically relevant perturbational model of planned myocardial infarction (PMI), patients undergoing septal ablation for hypertrophic cardiomyopathy. Blood samples were obtained before and at 10 and 60 minutes after PMI, and protein changes were assessed by repeated-measures analysis of variance. The generalizability of our PMI findings was evaluated in a spontaneous myocardial infarction cohort (Wilcoxon rank-sum). We then tested the platform's ability to detect associations between proteins and Framingham Risk Score components in the Framingham Heart Study, performing regression analyses for each protein versus each clinical trait.
  
  
  
  We found 217 proteins that significantly changed in the peripheral vein blood after PMI in a derivation cohort (n=15; P<5.70E-5). Seventy-nine of these proteins were validated in an independent PMI cohort (n=15; P<2.30E-4); >85% were directionally consistent and reached nominal significance. We detected many protein changes that are novel in the context of myocardial injury, including Dickkopf-related protein 4, a WNT pathway inhibitor (peak increase 124%, P=1.29E-15) and cripto, a growth factor important in cardiac development (peak increase 64%, P=1.74E-4). Among the 40 validated proteins that increased within 1 hour after PMI, 23 were also elevated in patients with spontaneous myocardial infarction (n=46; P<0.05). Framingham Heart Study analyses revealed 156 significant protein associations with the Framingham Risk Score (n=899), including aminoacylase 1 (β=0.3386, P=2.54E-22) and trigger factor 2 (β=0.2846, P=5.71E-17). Furthermore, we developed a novel workflow integrating DNA-based immunoaffinity with mass spectrometry to analytically validate aptamer specificity.
  
  
  
  Our results highlight an emerging proteomics tool capable of profiling >1000 low-abundance analytes with high sensitivity and high precision, applicable both to well-phenotyped perturbational studies and large human cohorts, as well.
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Debby
authors.initials: D
authors.lastname: Ngo
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Sumita
authors.initials: S
authors.lastname: Sinha
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Dongxiao
authors.initials: D
authors.lastname: Shen
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Eric W
authors.initials: EW
authors.lastname: Kuhn
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Michelle J
authors.initials: MJ
authors.lastname: Keyes
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Xu
authors.initials: X
authors.lastname: Shi
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Mark D
authors.initials: MD
authors.lastname: Benson
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: John F
authors.initials: JF
authors.lastname: O'Sullivan
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Hasmik
authors.initials: H
authors.lastname: Keshishian
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Laurie A
authors.initials: LA
authors.lastname: Farrell
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Michael A
authors.initials: MA
authors.lastname: Fifer
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Ramachandran S
authors.initials: RS
authors.lastname: Vasan
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Marc S
authors.initials: MS
authors.lastname: Sabatine
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Martin G
authors.initials: MG
authors.lastname: Larson
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.). rgerszte@bidmc.harvard.edu scarr@broadinstitute.org.
authors.forename: Steven A
authors.initials: SA
authors.lastname: Carr
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.). rgerszte@bidmc.harvard.edu scarr@broadinstitute.org.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Wang
authors.affiliation: From Division of Pulmonary and Critical Care Medicine, Department of Medicine (D.N.) and the Cardiovascular Research Center (D.N., S.S., D.S., M.J.K., X.S., M.D.B., J.F.O., L.A.F., R.E.G.), Massachusetts General Hospital, Harvard Medical School, Boston; Broad Institute of MIT and Harvard, Cambridge, MA (E.W.K., H.K., S.A.C., R.E.G.); Cardiovascular Division, Brigham and Women's Hospital, Boston, MA (M.D.B., M.S.S.); Cardiology Division, Massachusetts General Hospital, Harvard Medical School, Boston (M.A.F., M.S.S., R.E.G.); Preventive Medicine Section, Department of Medicine, Boston University School of Medicine, MA (R.S.V.); Department of Biostatistics, Boston University School of Public Health, MA (M.G.L.); The National Heart, Lung, and Blood Institute's Framingham Heart Study, Framingham, MA (M.G.L.); Division of Cardiovascular Medicine, Vanderbilt University, Nashville, TN (T.J.W.); and Division of Cardiovascular Medicine, Beth Israel Deaconess Medical Center, Boston, MA (R.E.G.).
authors.forename: Robert E
authors.initials: RE
authors.lastname: Gerszten
mesh_terms: D054058:Acute Coronary Syndrome / Q000097:blood; D000328:Adult; D052157:Aptamers, Nucleotide / Q000378:metabolism*; D015415:Biomarkers / Q000097:blood*; D001798:Blood Proteins / Q000032:analysis*; D002312:Cardiomyopathy, Hypertrophic / Q000097:blood* / Q000150:complications; D002853:Chromatography, Liquid; D004277:DNA, Single-Stranded / Q000378:metabolism; D005260:Female; D006801:Humans; D008137:Longitudinal Studies; D008297:Male; D008875:Middle Aged; D009203:Myocardial Infarction / Q000097:blood* / Q000209:etiology; D010641:Phenotype; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D053719:Tandem Mass Spectrometry
chemical_list: D052157:Aptamers, Nucleotide; D015415:Biomarkers; D001798:Blood Proteins; D004277:DNA, Single-Stranded
keywords: aptamer, nucleotides; cardiovascular diseases; mass spectrometry; myocardial infarction; proteomics