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Expression of Cyclic AMP associated genes on primary culture human myometrial cells

ABSTRACT: 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 ÂµM) for 48 hours, and then total RNA were extracted from each culture. Two comparisons were carried out including: 1. Control 2. Forksolin

ORGANISM(S): Homo sapiens

SUBMITTER: Suren Sooranna

PROVIDER: E-GEOD-75799 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Expression of Cyclic AMP associated genes on primary culture human myometrial cells

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.

2016-07-01 | GSE75799 | GEO

Identification of a Myometrial Molecular Profile for Dystocic Labor

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 Women in spontaneous labour undergoing cesarean section for dystocia (slow progressing labour) compared to women who had progressed in the second stage

2011-09-16 | E-GEOD-32178 | biostudies-arrayexpress

Genome-wide analysis of Prokineticin-1-regulated genes in human term myometrium

Project description:Infiltration of human myometrium and cervix with leukocytes and formation of a pro-inflammatory environment within the uterus has been associated with the initiation of both term and preterm parturition. The mechanism regulating the onset of this pro-inflammatory cascade is not fully elucidated. We demonstrate that prokineticin 1 (PROK1) is up-regulated in human myometrium and placenta during labour. Gene array analysis identified 65 genes up-regulated by PROK1 in human myometrium, mainly cytokines and chemokines including: Interleukin-1beta (IL1B), chemokine C-C motif ligand 3 (CCL3) and Colony Stimulating Factor 3 (CSF3). We additionally demonstrate that PROK1 increases expression of chemokines C-C motif ligand 20 (CCL20), Interleukin-6 (IL-6), Interleukin-8 (IL-8), prostaglandin synthase 2 (PTGS2) and prostaglandin E2 and F2? secretion. We propose that PROK1 is a novel inflammatory mediator that can contribute to onset of human parturition at term and partially mediate premature onset of inflammatory pathways during bacterial infection. Total RNA was extracted from human term non-labour myometrium explants treated with prokineticin for 6 and 24 hours compared to vehicle-treated explants. Six biological replicates were analyzed for each treatment and time point. Two of the 6-hour vehicle samples failed a quality-control analysis and were substituted with a 4-hour vehicle treatment from the same tissue sample in each case.

2011-12-31 | E-GEOD-28272 | biostudies-arrayexpress

RNA-seq Analysis of Changes in mRNA Expression for the hTERT-HMA/B Human Myometrial Cell Line treated with all Combinations of Progesterone, Forskolin, and Interleukin-1β

Project description:Parturition involves cellular signaling changes driven by the complex interplay between progesterone (P4), inflammation, and the cyclic adenosine monophosphate (cAMP) pathway. To characterize this interplay, comprehensive transcriptomic studies utilizing 8 treatment combinations were performed on the hTERT-HMA/B telomerase immortalized human myometrial cell line and myometrium obtained from term c-section deliveries. Genome-wide RNA-sequencing was performed on total RNA extracted from hTERT-HMA/B cells treated with all combinations of P4, forskolin (induces cAMP), and interleukin-1β (IL-1β). Gene set enrichment and regulatory network analyses was then used to identify pathways commonly, differentially, or synergistically regulated by each treatment. Tissue similarity index (TSI) was also applied to characterize the correspondence between cell lines and tissue phenotypes. In addition to their individual anti-inflammatory effects, P4 and cAMP synergistically blocked specific inflammatory pathways/regulators including STAT3/6, CEBPA/B, and OCT1/7, but not NFKB. TSI analysis indicated that forskolin+P4- and IL-1β-treated cells exhibit transcriptional signatures highly similar to non-laboring and laboring term myometrium, respectively. The data identify potential therapeutic targets to prevent preterm birth and show that the hTERT-HMA/B cell line provides is an accurate transcriptional model for term ± labor myometrium.

2019-07-26 | GSE134896 | GEO

Transcriptomic profiles of nonpregnant and term pregnant human myometrial tissues

Project description:The myometrium undergoes structural and functional remodeling during pregnancy. We hypothesize that myometrial genomic elements alter correspondingly in preparation for parturition. Human myometrial tissues from nonpregnant (NP) and term pregnant (TP) human subjects were subject to RNAseq, ATAC-seq and PGR ChIP-seq assays to profile transcriptome, assessible genome and PGR occupancy.

2019-11-28 | GSE137551 | GEO

Comparison of the myometrial transcriptome from singleton and twin pregnancies by high throughput RNA-Seq

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.

2020-01-08 | E-MTAB-8233 | biostudies-arrayexpress

Identification of a Myometrial Molecular Profile for Dystocic Labor

2011-09-17 | GSE32178 | GEO

RNA-seq transcriptomes of term not in labour and term in labour human myometrial tissue

Project description:Purpose: To chart the human myometrial transcriptomes before and after the onset of labour. Methods: Tophat splice junction mapping of paired-end reads, HTSeq to generate counts, cufflinks to track transcripts, DESeq, edgeR and baySeq to detect differentially expressed genes and principal component analysis for clustering analyses. Results: We mapped on average 14 million paired-end reads per sample (counting each end individually) to the human genome (build hg19) and covered the expressed transcriptome about 13 times with a TopHat-HTSeq workflow. We performed a comparative analysis with an analogous microarray study (Mittal et al., 2010) and found some overlap between the RNA-seq and the microarray data. Conclusions: Our study is the first RNA-seq study of the human myometrium before and after the onset of labour. We show that while microarray and RNA-seq studies may complement each other, RNA-seq has a much greater resolution. At term with and at term without labour human myometrial mRNA profiles were generated by deep sequencing, using Illumina GAIIx (five biological replicates each).

2014-02-17 | E-GEOD-50599 | biostudies-arrayexpress

MeDIP assay using human myometrium of women with preterm labor of different phenotypes vs. normal term labor

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

2017-05-04 | E-GEOD-71123 | biostudies-arrayexpress

Comparison of the in vivo transcriptome of the myometrium to the transcriptome of myometrial explants, primary cultured myometrial cells and the hTert myometrial cell line

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.

2016-05-30 | GSE77830 | GEO

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