Identification of a Myometrial Molecular Profile for Dystocic Labor
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ABSTRACT: This study identifies a transciptomic myometrial profile associated with dystocia in spontanous nulliparous term labour We used microarrays to compare myometrial biopsies obtained at cesarean section from women in spontaneous term labour Women in spontaneous labour undergoing cesarean section for dystocia (slow progressing labour) compared to women who had progressed in the second stage
Project description:This study identifies a transciptomic myometrial profile associated with dystocia in spontanous nulliparous term labour We used microarrays to compare myometrial biopsies obtained at cesarean section from women in spontaneous term labour
Project description:During pregnancy, the myometrium remains quiescent but at term, switches to a state capable of producing a series of coordinated contractions for the delivery of the fetus. Myometrial contractions of labour signify the normal physiological end-point of pregnancy but the biochemical onset of labour may occur at or before term via a series of changes in expression of labour associated genes that are responsible for controlling the activity of the uterus during pregnancy and parturition. There is increasing evidence that components of the cAMP-signalling pathway are up-regulated in the human myometrium during pregnancy to promote the relaxation of the myometrium until term. Our aim was to determine which cAMP-associated genes are important during pregnancy and parturition by exposing myometrial cells to forskolin and performing an a gene array. We then plan to study the trend of the cAMP-associated genes at different stages of gestation and during labour. In this study, we used microarrays to elucidate forskolin responsive genes in human myometrium. These data may provide a broader view of gene networks and cellular functions regulated by forskolin in human myometrial cells. In our future study, this will also help us understand the role of cAMP in human parturition. Primary cultures of human myometrial cells were grown from myometrial biopsies obtained at the time of elective caesarean section at term. Cells were exposed to forskolin (100 µM) for 48 hours, and then total RNA were extracted from each culture. Two comparisons were carried out including: 1. Control 2. Forksolin
Project description:Differentially expressed mRNA transcripts in the placenta delivered by term spontaneous labour compared to those delivered by elective term cesarean section. We hypothesized that the labour process involves changes in mRNA expression in the placenta. To test this hypothesis, we interrogated the mRNA levels of >50,000 genes and transcript variants using gene expression microarray (Human Genome U133 Plus 2.0 Array, Affymetrix) on 5 placentas collected from term spontaneous delivery and another 5 placentas collected from elective term cesarean delivery. To minimize the effect of gestational age on gene expression, these two groups of placentas were matched for their gestational ages at delivery. We have identified 134 and 128 genes that were up- or down-regulated, respectively, for more than 3-fold in the term (spontaneous) labour placentas compared to the term (elective) cesarean placentas (Mann-Whitney Rank Sum Test, p-value <= 0.05 after Benjamini and Hochberg adjustment for multiple testing). Experiment Overall Design: Placentas collected from (i) term spontaeous labour and (ii) elective term cesarean section were subjected to RNA extraction and hybridization on Affymetrix microarrays. To identify gene expression patterns that are commonly involved in term spontaneous labour, we analyzed 5 placentas from each of these 2 groups and tested for any differentially expressed genes by non-parametric statistical methods.
Project description:Circulating progesterone (P4) levels decline before the onset of parturition in most animals, but not in humans. This has led to the suggestion that there is functional withdrawal of P4 action at the myometrial level prior to labor onset. Mifepristone is widely used to induce human labour In this study, we aimed to establish and validate a model of human myometrial explants for the study of P4 action. Myometrial biopsies obtained at Caesearean section at term were dissected into explants after a portion was immediately snap-frozen (t=0). Transcriptomic comparison of paired explants and primary myometrial cells as well as the hTert immortalized myometrial cell line demonstrated that explants more closely resemble t=0. Biopsies obtained from non-laboring women at elective Caesarean section at term were divided into 3: (i) dissected and immediately snap-frozen (t=0), (ii) dissected into 3x3x3mm3myometrial explants and (iii) processed for primary cell culture. Explants, primary cells at passage 4 (the typical passage our group uses for experiments) and hTERT cells were cultured for a period of 30 hours without treatment. Total RNA was extracted and microarray analysis performed. 6 replicates were used for this study.
Project description:Differentially expressed mRNA transcripts in the placenta delivered by term spontaneous labour compared to those delivered by elective term cesarean section. We hypothesized that the labour process involves changes in mRNA expression in the placenta. To test this hypothesis, we interrogated the mRNA levels of >50,000 genes and transcript variants using gene expression microarray (Human Genome U133 Plus 2.0 Array, Affymetrix) on 5 placentas collected from term spontaneous delivery and another 5 placentas collected from elective term cesarean delivery. To minimize the effect of gestational age on gene expression, these two groups of placentas were matched for their gestational ages at delivery. We have identified 134 and 128 genes that were up- or down-regulated, respectively, for more than 3-fold in the term (spontaneous) labour placentas compared to the term (elective) cesarean placentas (Mann-Whitney Rank Sum Test, p-value <= 0.05 after Benjamini and Hochberg adjustment for multiple testing).
Project description:We performed high-throughput RNA sequencing (RNA-Seq) of human myometrium from singleton and twin pregnancies at term (> 37+0 weeks) and preterm (< 37+0 weeks), collected during pre-labour Caesarean Section.
Project description:Preterm birth is multifactorial in origin with several distinct clinical phenotypes of differing etiologies, including idiopathic preterm birth. Preterm birth involves the interaction of genetic, societal and environmental factors such as nutrition, lifestyle and stress that may modulate the length of gestation via the epigenome. DNA methylation is a well-studied epigenetic modification whereby promoter methylation commonly represses gene expression and vice versa. Myometrial tissue was obtained at cesarean section at term with or without labor, preterm without labor, idiopathic preterm labor, and twin gestations with labor. Differences in the myometrial epigenomes were identified at gene promoters, CpG islands, CpG island shores and shelves, gene bodies across the genome between the groups of women with preterm labor of different phenotypes vs. normal term labor. Functional clustering analysis indicated the significantly enriched pathways of hypomethylated genes (permissive) were related to acute inflammatory and acute-phase responses. By contrast, genes that are hypermethylated (repressive) revealed enrichment for contractile fibers and cell. This study provides the first high-resolution DNA methylome of human myometrium with evidence for differences in the methylome that may relate to idiopathic preterm birth via regulation of gene expression. The findings extend previous observations that idiopathic preterm labor is associated with subclinical intrauterine infection and inflammatory pathways and point to targets for further molecular characterization of preterm delivery. Comparison of the human myometrial epigenomes in pregnancies with preterm labor of different phenotypes vs. normal term labor
Project description:Circulating progesterone (P4) levels decline before the onset of parturition in most animals, but not in humans. This has led to the suggestion that there is functional withdrawal of P4 action at the myometrial level prior to labor onset. Mifepristone is widely used to induce human labour In this study, we aimed to establish and validate a model of human myometrial explants for the study of P4 action. Myometrial biopsies obtained at Caesearean section at term were dissected into explants after a portion was immediately snap-frozen (t=0). Transcriptomic comparison of paired explants and primary myometrial cells as well as the hTert immortalized myometrial cell line demonstrated that explants more closely resemble t=0.
Project description:Myometrial biopsies were collected from 20 women undergoing primary cesarean sections in well-characterized clinical scenarios: 1) term labor of spontaneous onset (TL, n=5); 2) term non-labor (TNL, n=5); 3) spontaneous PTB in the setting of chorioamnionitis (PTB-HCA) and 4) indicated preterm birth (PTB) non-labor (PTB-NL, n=5). RNAs were profiled using 2nd-generation RNA sequencing.
Project description:Myometrial biopsies were collected from 31 women undergoing primary cesarean sections and were carefully phenotyped with respect to gestational age (GA), circumstances of labor onset, and clinical status at the start and end of the intervention. Cases were aggregated into groups as follows: Group 1: term birth following spontaneous onset of term labor (TL, n=5); Group 2: term birth by elective cesarean section not in labor (TNL, n=5); Group 3: PTB following spontaneous preterm labor with intact membranes (n=6); Group 4: preterm birth following PPROM (n=8); and Group 5: provider-initiated preterm birth in the absence of active labor contractions, cervical dilation or membrane rupture (n=7). Additional phenotyping of cases spontaneously committed to PTBs (Groups 3 and 4) involved presence or absence of Triple I based on cultures of amniotic fluid obtained via clinically-indicated amniocentesis. RNAs were profiled using second generation RNA sequencing.