Isobaric labeling and tandem mass spectrometry: A novel approach for profiling and quantifying proteins differentially expressed in amniotic fluid in preterm labor with and without intra-amniotic infection/inflammation

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dc.contributor.authorRomero, Roberto
dc.contributor.authorKusanovic, Juan Pedro
dc.contributor.authorGotsch, Francesca
dc.contributor.authorErez, Offer
dc.contributor.authorVaisbuch, Edi
dc.contributor.authorMazaki Tovi, Shali
dc.contributor.authorMoser, Allan
dc.contributor.authorTam, Sunny
dc.contributor.authorLeszyk, John
dc.contributor.authorMaster, Stephen R.
dc.contributor.authorJuhasz, Peter
dc.contributor.authorPacora, Percy
dc.contributor.authorOgge, Giovanna
dc.contributor.authorGomez, Ricardo
dc.contributor.authorYoon, Bo H.
dc.contributor.authorYeo, Lami
dc.contributor.authorHassan, Sonia S.
dc.contributor.authorRogers, Wade T.
dc.date.accessioned2024-01-10T13:49:41Z
dc.date.available2024-01-10T13:49:41Z
dc.date.issued2010
dc.description.abstractMethods. A cross-sectional study was designed and included AF samples from patients with spontaneous PTL and intact membranes in the following groups: (1) patients without IAI who delivered at term (n = 26); (2) patients who delivered preterm without IAI (n = 25); and (3) patients with IAI (n = 24). Proteomic profiling of AF samples was performed using a workflow involving tryptic digestion, iTRAQ labeling and multiplexing, strong cation exchange fractionation, and liquid chromatography tandem mass spectrometry. Twenty-five separate 4-plex samples were prepared and analyzed.
dc.description.abstractResults. Collectively, 123,011 MS<SU2</SU spectra were analyzed, and over 25,000 peptides were analyzed by database search (X!Tandem and Mascot), resulting in the identification of 309 unique high-confidence proteins. Analysis of differentially present iTRAQ reporter peaks revealed many proteins that have been previously reported to be associated with preterm delivery with IAI. Importantly, many novel proteins were found to be up-regulated in the AF of patients with PTL and IAI including leukocyte elastase precursor, Thymosin-like 3, and 14-3-3 protein isoforms. Moreover, we observed differential expression of proteins in AF of patients who delivered preterm in the absence of IAI in comparison with those with PTL who delivered at term including Mimecan precursor, latent-transforming growth factor beta-binding protein isoform 1L precursor, and Resistin. These findings have been confirmed for Resistin in an independent cohort of samples using ELISA. Gene ontology enrichment analysis was employed to reveal families of proteins participating in distinct biological processes. We identified enrichment for host defense, anti-apoptosis, metabolism/catabolism and cell and protein mobility, localization and targeting.
dc.description.abstractConclusions. (1) Proteomics with iTRAQ labeling is a profiling tool capable of revealing differential expression of proteins in AF; (2) We discovered 82 proteins differentially expressed in three clinical subgroups of premature labor, 67 which were heretofore unknown. Of particular importance is the identification of proteins differentially expressed in AF from women who delivered preterm in the absence of IAI. This is the first report of the positive identification of biomarkers in this subgroup of patients.
dc.description.funderPerinatology Research Branch, Division of Intramural Research, Eunice Kennedy Shriver National Institute of Child Health and Human Development, NIH, DHHS
dc.description.funderEUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
dc.description.funderEUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH &HUMAN DEVELOPMENT
dc.fechaingreso.objetodigital2024-05-23
dc.format.extent20 páginas
dc.fuente.origenWOS
dc.identifier.doi10.3109/14767050903067386
dc.identifier.eissn1476-4954
dc.identifier.issn1476-7058
dc.identifier.pubmedidMEDLINE:19670042
dc.identifier.urihttps://doi.org/10.3109/14767050903067386
dc.identifier.urihttps://repositorio.uc.cl/handle/11534/79472
dc.identifier.wosidWOS:000275470600003
dc.information.autorucMedicina;Gómez R;S/I;80926
dc.issue.numero4
dc.language.isoen
dc.nota.accesocontenido parcial
dc.pagina.final280
dc.pagina.inicio261
dc.publisherTAYLOR & FRANCIS LTD
dc.revistaJOURNAL OF MATERNAL-FETAL & NEONATAL MEDICINE
dc.rightsacceso restringido
dc.subjectProteomic
dc.subjectpreterm birth
dc.subjectprematurity
dc.subjectMIAC
dc.subjectmicrobial invasion of the amniotic cavity
dc.subjectchorioamnionitis
dc.subjectisotopic
dc.subjectmultiplex
dc.subjectlabel
dc.subjecthigh-dimensional biology
dc.subject'bottom-up'
dc.subject'top-down'
dc.subjectcomputational biology
dc.subjecthigh-dimensional biology
dc.subjectsystems biology
dc.subjectendoplasmic reticulum stress
dc.subjectmisfolded protein
dc.subjectunfolded protein
dc.subjectBLOOD-CELL COUNT
dc.subjectC-REACTIVE PROTEIN
dc.subjectMULTIDIMENSIONAL LIQUID-CHROMATOGRAPHY
dc.subjectACTIVATING PEPTIDE-1 INTERLEUKIN-8
dc.subjectTOLL-LIKE RECEPTORS
dc.subjectMICROBIAL INVASION
dc.subjectGENE-EXPRESSION
dc.subjectPREMATURE RUPTURE
dc.subjectGRAM STAIN
dc.subjectHISTOLOGIC CHORIOAMNIONITIS
dc.subject.ods05 Gender Equality
dc.subject.ods03 Good Health and Well-being
dc.subject.odspa05 Igualdad de género
dc.subject.odspa03 Salud y bienestar
dc.titleIsobaric labeling and tandem mass spectrometry: A novel approach for profiling and quantifying proteins differentially expressed in amniotic fluid in preterm labor with and without intra-amniotic infection/inflammation
dc.typeartículo
dc.volumen23
sipa.codpersvinculados80926
sipa.indexWOS
sipa.indexScopus
sipa.trazabilidadCarga SIPA;09-01-2024
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