732 AN IN-DEPTH ANALYSIS OF THE PEDIATRIC URINARY PROTEOME

Abstract

You have accessJournal of UrologyPediatrics: Basic Research1 Apr 2012732 AN IN-DEPTH ANALYSIS OF THE PEDIATRIC URINARY PROTEOME John W. Froehlich, Ali R. Vaezzadeh, Marc Kirchner, Andrew C. Briscoe, Oliver Hofmann, Winston Hide, Hanno Steen, and Richard S. Lee John W. FroehlichJohn W. Froehlich Boston, MA More articles by this author , Ali R. VaezzadehAli R. Vaezzadeh Boston, MA More articles by this author , Marc KirchnerMarc Kirchner Boston, MA More articles by this author , Andrew C. BriscoeAndrew C. Briscoe Boston, MA More articles by this author , Oliver HofmannOliver Hofmann Boston, MA More articles by this author , Winston HideWinston Hide Boston, MA More articles by this author , Hanno SteenHanno Steen Boston, MA More articles by this author , and Richard S. LeeRichard S. Lee Boston, MA More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2012.02.817AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Urine is a valuable body fluid for clinical analysis, and may be obtained noninvasively in large amounts. Many researchers are currently using urinary proteomics to identify biomarkers for specific diseases. Despite efforts to identify biomarkers in diseased samples, detailed proteomic analysis of non-diseased pediatric urine samples is still lagging behind. Furthermore, although age-specific changes are known to occur in rats, similar variations between pediatric and adult urines have not been determined. This study compares healthy male adult and infant urines, to establish which proteins are enriched or specific in infant's urine, and which proteins are present independent of age. METHODS Urine samples and demographic data were obtained from a urine specimen registry using an IRB approved protocol. Cellular debris was removed and 100 microgram protein aliquots were separated by PAGE. Proteins were digested with trypsin, extracted from the gel and purified before analysis by mass spectrometry using an LTQ-FT mass spectrometer (Thermo). MS data were searched against the uniprot KB database (homo sapiens). Peptides were filtered to ensure a 1% false discovery rate. A spectral counting approach was employed, using normalized spectral counts to minimize interpersonal variability. Statistically significant proteins (p<0.05) were identified, and analyzed by genome ontology to find overrepresented terms (p<0.0001) associated exclusively with adult and infant urines. RESULTS A total of 672 proteins were commonly identified in both cohorts, out of 1321 total protein identifications. Between our data and contemporary adult proteomic studies, 222 proteins were identified in infants alone. The infant-specific proteome was overrepresented in cellular metabolic processes, gene expression, and RNA processing. In addition, among commonly identified proteins, several proteins associated with glycolysis and ROS were enriched in the infant. These include pyruvate kinase isozymes M1/M2, Phosphoglycerate kinase 1, Aldose-1 epimerase, Retinal dehydrogenase 1, Fructose-1,6-bisphosphatase 1 and L-lactate dehydrogenase B chain. CONCLUSIONS We demonstrated that the male urinary proteome is highly age dependent, and hypothesize that the differences reflect differing growth and metabolic states between the adult and infant cohorts. These results highlight the importance of understanding the variation among non-diseased samples to better understand normal progression of the urinary proteome. © 2012 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 187Issue 4SApril 2012Page: e300-e301 Advertisement Copyright & Permissions© 2012 by American Urological Association Education and Research, Inc.MetricsAuthor Information John W. Froehlich Boston, MA More articles by this author Ali R. Vaezzadeh Boston, MA More articles by this author Marc Kirchner Boston, MA More articles by this author Andrew C. Briscoe Boston, MA More articles by this author Oliver Hofmann Boston, MA More articles by this author Winston Hide Boston, MA More articles by this author Hanno Steen Boston, MA More articles by this author Richard S. Lee Boston, MA More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...