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Multidimensional proteomics analysis of amniotic fluid to provide insight into the mechanisms of idiopathic preterm birth.

Buhimschi IA, Zhao G, Rosenberg VA, Abdel-Razeq S, Thung S, Buhimschi CS - PLoS ONE (2008)

Bottom Line: Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40) were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results.Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry) coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER) ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.The altered proteins may offer opportunities for therapeutical intervention and future drug development to prevent prematurity.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, Connecticut, United States of America. irina.buhimschi@yale.edu

ABSTRACT

Background: Though recent advancement in proteomics has provided a novel perspective on several distinct pathogenetic mechanisms leading to preterm birth (inflammation, bleeding), the etiology of most preterm births still remains elusive. We conducted a multidimensional proteomic analysis of the amniotic fluid to identify pathways related to preterm birth in the absence of inflammation or bleeding.

Methodology/principal findings: A proteomic fingerprint was generated from fresh amniotic fluid using surface-enhanced laser desorbtion ionization time of flight (SELDI-TOF) mass spectrometry in a total of 286 consecutive samples retrieved from women who presented with signs or symptoms of preterm labor or preterm premature rupture of the membranes. Inflammation and/or bleeding proteomic patterns were detected in 32% (92/286) of the SELDI tracings. In the remaining tracings, a hierarchical algorithm was applied based on descriptors quantifying similarity/dissimilarity among proteomic fingerprints. This allowed identification of a novel profile (Q-profile) based on the presence of 5 SELDI peaks in the 10-12.5 kDa mass area. Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40) were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results. Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry) coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER) ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.

Conclusion/significance: Proteomic profiling of amniotic fluid coupled with non-hierarchical bioinformatics algorithms identified a subgroup of patients at risk for preterm birth in the absence of intra-amniotic inflammation or bleeding, suggesting a novel pathogenetic pathway leading to preterm birth. The altered proteins may offer opportunities for therapeutical intervention and future drug development to prevent prematurity.

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Representative two color merged 2D-DIGE image with up or down-regulated proteins selected for identification.Proteins in amniotic fluid with Q-profile present were labeled with Cy3 and proteins in control fluids with Cy5. Spots differentially regulated (≥1.5 fold in log spot volume) were selected for identification. Spots up-regulated are circled in red and spots down-regulated are circled in blue.
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pone-0002049-g004: Representative two color merged 2D-DIGE image with up or down-regulated proteins selected for identification.Proteins in amniotic fluid with Q-profile present were labeled with Cy3 and proteins in control fluids with Cy5. Spots differentially regulated (≥1.5 fold in log spot volume) were selected for identification. Spots up-regulated are circled in red and spots down-regulated are circled in blue.

Mentions: Figure 4 shows a representative 2D-DIGE image of proteins differentially expressed (red: spots up-regulated ≥1.5-fold, blue: spots down-regulated≥1.5-fold) between samples displaying the Q-profile and reference samples amniotic fluid samples (matched for gestational age at time of amniocentesis). Interestingly, more spots (n = 60) appeared down-regulated in the amniotic fluids displaying the Q-profile rather than up-regulated (n = 28). Of the spots picked for identification, 22 matched to a protein in the database with more than one peptide. Few spots matched to the same NCBI GenID transcript number. In fact, a total of 17 distinct protein database matches appeared and were differentially regulated at least 1.5-fold (Table 4) among the 2 groups. Further data mining using the PANTHER classification system revealed that of the unambiguous identities, 10 protein products belonged to 7 distinct and well-classified biological processes. The most populated was protein metabolism (5/10 transcripts matched) and signal transduction (3/10 transcripts matched) (Figure 5). PANTHER further converged the aberrantly expressed identities by matching several transcripts to the same protein precursor in the database (Table 5). This allowed us to conclude that the amniotic fluids with Q-profile are characterized by up-regulation in insulin growth factor binding protein (IGFBP-1), apolipoproteins (APO A-I and APO A-IV), lumican and bikunin (alpha-1-microglobulin light chain protein, complex-forming glycoprotein heterogeneous in charge) and down-regulation in the anti-proteolytic factors alpha-1-antitrypsin (SERPINA1), alpha-2-antiplasmin (SEPRPINF1, pigment epithelium derived factor) (Table 5).


Multidimensional proteomics analysis of amniotic fluid to provide insight into the mechanisms of idiopathic preterm birth.

Buhimschi IA, Zhao G, Rosenberg VA, Abdel-Razeq S, Thung S, Buhimschi CS - PLoS ONE (2008)

Representative two color merged 2D-DIGE image with up or down-regulated proteins selected for identification.Proteins in amniotic fluid with Q-profile present were labeled with Cy3 and proteins in control fluids with Cy5. Spots differentially regulated (≥1.5 fold in log spot volume) were selected for identification. Spots up-regulated are circled in red and spots down-regulated are circled in blue.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2315798&req=5

pone-0002049-g004: Representative two color merged 2D-DIGE image with up or down-regulated proteins selected for identification.Proteins in amniotic fluid with Q-profile present were labeled with Cy3 and proteins in control fluids with Cy5. Spots differentially regulated (≥1.5 fold in log spot volume) were selected for identification. Spots up-regulated are circled in red and spots down-regulated are circled in blue.
Mentions: Figure 4 shows a representative 2D-DIGE image of proteins differentially expressed (red: spots up-regulated ≥1.5-fold, blue: spots down-regulated≥1.5-fold) between samples displaying the Q-profile and reference samples amniotic fluid samples (matched for gestational age at time of amniocentesis). Interestingly, more spots (n = 60) appeared down-regulated in the amniotic fluids displaying the Q-profile rather than up-regulated (n = 28). Of the spots picked for identification, 22 matched to a protein in the database with more than one peptide. Few spots matched to the same NCBI GenID transcript number. In fact, a total of 17 distinct protein database matches appeared and were differentially regulated at least 1.5-fold (Table 4) among the 2 groups. Further data mining using the PANTHER classification system revealed that of the unambiguous identities, 10 protein products belonged to 7 distinct and well-classified biological processes. The most populated was protein metabolism (5/10 transcripts matched) and signal transduction (3/10 transcripts matched) (Figure 5). PANTHER further converged the aberrantly expressed identities by matching several transcripts to the same protein precursor in the database (Table 5). This allowed us to conclude that the amniotic fluids with Q-profile are characterized by up-regulation in insulin growth factor binding protein (IGFBP-1), apolipoproteins (APO A-I and APO A-IV), lumican and bikunin (alpha-1-microglobulin light chain protein, complex-forming glycoprotein heterogeneous in charge) and down-regulation in the anti-proteolytic factors alpha-1-antitrypsin (SERPINA1), alpha-2-antiplasmin (SEPRPINF1, pigment epithelium derived factor) (Table 5).

Bottom Line: Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40) were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results.Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry) coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER) ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.The altered proteins may offer opportunities for therapeutical intervention and future drug development to prevent prematurity.

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics, Gynecology and Reproductive Sciences, Yale University School of Medicine, New Haven, Connecticut, United States of America. irina.buhimschi@yale.edu

ABSTRACT

Background: Though recent advancement in proteomics has provided a novel perspective on several distinct pathogenetic mechanisms leading to preterm birth (inflammation, bleeding), the etiology of most preterm births still remains elusive. We conducted a multidimensional proteomic analysis of the amniotic fluid to identify pathways related to preterm birth in the absence of inflammation or bleeding.

Methodology/principal findings: A proteomic fingerprint was generated from fresh amniotic fluid using surface-enhanced laser desorbtion ionization time of flight (SELDI-TOF) mass spectrometry in a total of 286 consecutive samples retrieved from women who presented with signs or symptoms of preterm labor or preterm premature rupture of the membranes. Inflammation and/or bleeding proteomic patterns were detected in 32% (92/286) of the SELDI tracings. In the remaining tracings, a hierarchical algorithm was applied based on descriptors quantifying similarity/dissimilarity among proteomic fingerprints. This allowed identification of a novel profile (Q-profile) based on the presence of 5 SELDI peaks in the 10-12.5 kDa mass area. Women displaying the Q-profile (mean+/-SD, gestational age: 25+/-4 weeks, n = 40) were more likely to deliver preterm despite expectant management in the context of intact membranes and normal amniotic fluid clinical results. Utilizing identification-centered proteomics techniques (fluorescence two-dimensional differential gel electrophoresis, robotic tryptic digestion and mass spectrometry) coupled with Protein ANalysis THrough Evolutionary Relationships (PANTHER) ontological classifications, we determined that in amniotic fluids with Q-profile the differentially expressed proteins are primarily involved in non-inflammatory biological processes such as protein metabolism, signal transduction and transport.

Conclusion/significance: Proteomic profiling of amniotic fluid coupled with non-hierarchical bioinformatics algorithms identified a subgroup of patients at risk for preterm birth in the absence of intra-amniotic inflammation or bleeding, suggesting a novel pathogenetic pathway leading to preterm birth. The altered proteins may offer opportunities for therapeutical intervention and future drug development to prevent prematurity.

Show MeSH
Related in: MedlinePlus