Reductionist and system approaches to study the role of infection in preterm labor and delivery

A substantial number of patients with preterm labor and delivery do not show clinical signs of infection, however, it is the subclinical form which is the main causative factor and often results in premature delivery. The hitherto commonly applied methods of inflammation detection are based either on potentially hazardous amniocentesis or still insufficient inflammation-related protein measurement in the serum or other biological fluids

Surfactant proteins SP-A and SP-D modulate uterine contractile events in ULTR myometrial cell line

Pulmonary surfactant proteins SP-A and SP-D are pattern recognition innate immune molecules. However, there is extrapulmonary existence, especially in the amniotic fluid and at the feto-maternal interface. There is sufficient evidence to suggest that SP-A and SP-D are involved in the initiation of labour. This is of great importance given that preterm birth is associated with increased mortality and morbidity. In this study, we investigated the effects of recombinant forms of SP-A and SP-D (rhSP-A and rhSP-D, the comprising of trimeric lectin domain) on contractile events in vitro, using a human myometrial cell line (ULTR) as an experimental model. Treatment with rhSP-A or rhSP-D increased the cell velocity, distance travelled and displacement by ULTR cells. rhSP-A and rhSP-D also affected the contractile response of ULTRs when grown on collagen matrices showing reduced surface area. We investigated this effect further by measuring contractility-associated protein (CAP) genes. Treatment with rhSP-A and rhSP-D induced expression of oxytocin receptor (OXTR) and connexin 43 (CX43). In addition, rhSP-A and rhSP-D were able to induce secretion of GROα and IL-8. rhSP-D also induced the expression of IL-6 and IL-6 Ra. We provide evidence that SP-A and SP-D play a key role in modulating events prior to labour by reconditioning the human myometrium and in inducing CAP genes and pro-inflammatory cytokines thus shifting the uterus from a quiescent state to a contractile one

Early pregnancy peripheral blood gene expression and risk of preterm delivery: a nested case control study

<p>Abstract</p> <p>Background</p> <p>Preterm delivery (PTD) is a significant public health problem associated with greater risk of mortality and morbidity in infants and mothers. Pathophysiologic processes that may lead to PTD start early in pregnancy. We investigated early pregnancy peripheral blood global gene expression and PTD risk.</p> <p>Methods</p> <p>As part of a prospective study, ribonucleic acid was extracted from blood samples (collected at 16 weeks gestational age) from 14 women who had PTD (cases) and 16 women who delivered at term (controls). Gene expressions were measured using the GeneChip^®Human Genome U133 Plus 2.0 Array. Student's T-test and fold change analysis were used to identify differentially expressed genes. We used hierarchical clustering and principle components analysis to characterize signature gene expression patterns among cases and controls. Pathway and promoter sequence analyses were used to investigate functions and functional relationships as well as regulatory regions of differentially expressed genes.</p> <p>Results</p> <p>A total of 209 genes, including potential candidate genes (e.g. PTGDS, prostaglandin D2 synthase 21 kDa), were differentially expressed. A set of these genes achieved accurate pre-diagnostic separation of cases and controls. These genes participate in functions related to immune system and inflammation, organ development, metabolism (lipid, carbohydrate and amino acid) and cell signaling. Binding sites of putative transcription factors such as EGR1 (early growth response 1), TFAP2A (transcription factor AP2A), Sp1 (specificity protein 1) and Sp3 (specificity protein 3) were over represented in promoter regions of differentially expressed genes. Real-time PCR confirmed microarray expression measurements of selected genes.</p> <p>Conclusions</p> <p>PTD is associated with maternal early pregnancy peripheral blood gene expression changes. Maternal early pregnancy peripheral blood gene expression patterns may be useful for better understanding of PTD pathophysiology and PTD risk prediction.</p

Transcription analysis of the myometrium of labouring and non-labouring women

An incomplete understanding of the molecular mechanisms that initiate normal human labour at term seriously hampers the development of effective ways to predict, prevent and treat disorders such as preterm labour. Appropriate analysis of large microarray experiments that compare gene expression in non-labouring and labouring gestational tissues is necessary to help bridge these gaps in our knowledge. In this work, gene expression in 48 (22 labouring, 26 non-labouring) lower-segment myometrial samples collected at Caesarean section were analysed using Illumina HT-12 v4.0 BeadChips. Normalised data were compared between labouring and non-labouring groups using traditional statistical methods and a novel network graph approach. We sought technical validation with quantitative real-time PCR, and biological replication through inverse variance-weighted meta-analysis with published microarray data. We have extended the list of genes suggested to be associated with labour: Compared to non-labouring samples, labouring samples showed apparent higher expression at 960 probes (949 genes) and apparent lower expression at 801 probes (789 genes) (absolute fold change ≥1.2, rank product percentage of false positive value (RP-PFP) <0.05). Although half of the women in the labouring group had received pharmaceutical treatment to induce or augment labour, sensitivity analysis suggested that this did not confound our results. In agreement with previous studies, functional analysis suggested that labour was characterised by an increase in the expression of inflammatory genes and network analysis suggested a strong neutrophil signature. Our analysis also suggested that labour is characterised by a decrease in the expression of muscle-specific processes, which has not been explicitly discussed previously. We validated these findings through the first formal meta-analysis of raw data from previous experiments and we hypothesise that this represents a change in the composition of myometrial tissue at labour. Further work will be necessary to reveal whether these results are solely due to leukocyte infiltration into the myometrium as a mechanism initiating labour, or in addition whether they also represent gene changes in the myocytes themselves. We have made all our data available at www.ebi.ac.uk/arrayexpress/ (accession number E-MTAB-3136) to facilitate progression of this work

ROLE DES ISOFORMES DE PROTEINE KINASE C DANS LE CONTROLE DE LA CROISSANCE ET DE L'ACTIVITE CONTRACTILE DU MUSCLE UTERIN HUMAIN. REGULATION PAR L'ENDOTHELINE-1 ET ETUDE PHYSIOPATHOLOGIQUE

DANS LE MUSCLE LISSE UTERIN HUMAIN, DEUX PROPRIETES DE L'ENDOTHELINE-1 (ET-1) ONT ETE MISES EN EVIDENCE. SA CAPACITE A INDUIRE RAPIDEMENT DES CONTRACTIONS UTERINES IMPLIQUE UN ROLE DE CE PEPTIDE DANS LA PARTURITION HUMAINE, ET SON EFFET MITOGENE A PLUS LONG TERME SUR LES CELLULES MYOMETRIALES SUGGERE UN ROLE DE ET-1 DANS LE DEVELOPPEMENT NORMAL ET PHYSIOPATHOLOGIQUE DU MYOMETRE. AU COURS DE CE TRAVAIL, NOUS AVONS CARACTERISE, DANS LE MYOMETRE HUMAIN, LE SYSTEME ENZYMATIQUE PROTEINE KINASE C (PKC), SITUE AU CUR DE MECANISMES PHYSIOLOGIQUES (CONTRACTION, PROLIFERATION, DIFFERENCIATION) ET PHYSIOPATHOLOGIQUES (TUMORIGENESE). NOUS AVONS RECHERCHE SON IMPLICATION DANS LA TRANSDUCTION DU SIGNAL CONTRACTILE ET MITOGENE DE ET-1 DANS LE MYOMETRE NON GESTANT ET GESTANT. NOUS AVONS ETENDU NOTRE ETUDE AUX LEIOMYOMES UTERINS, QUI CONSTITUENT UN EXEMPLE DE DEVELOPPEMENT ANARCHIQUE DES CELLULES MYOMETRIALES. L'IDENTIFICATION DU SYSTEME PKC NOUS A PERMIS DE SOULIGNER QUE LES SIX ISOFORMES DE PKC (, 1, 2, , ET ) DETECTEES DANS LE MYOMETRE ISSU DE FEMMES NON GESTANTES SONT EGALEMENT PRESENTES DANS LE MYOMETRE DE FEMMES GESTANTES A TERME, DANS LES CELLULES MYOMETRIALES HUMAINES EN CULTURE, DANS LES LEIOMYOMES ET DANS LES CELLULES DE LEIOMYOMES HUMAINES. LA REGULATION DIFFERENTIELLE DES ISOFORMES DE PKC SOUS L'ACTION DE ET-1 DANS CES DIFFERENTES SITUATIONS SUGGERE QUE L'ACTIVITE CONTRACTILE ET LA CROISSANCE CELLULAIRE SONT, DANS LE MUSCLE LISSE UTERIN, REGULEES PAR DES ISOFORMES DE PKC SPECIFIQUES. DANS LE MYOMETRE DE FEMMES GESTANTES A TERME, NOUS AVONS MIS EN EVIDENCE UN ROLE DETERMINANT DES ISOFORMES DE PKC ET/OU DANS L'ACTION CONTRACTILE DE ET-1. DANS LES CELLULES MYOMETRIALES HUMAINES EN CULTURE, NOUS AVONS MONTRE QUE LA PKC EST RESPONSABLE DE L'EFFET MITOGENE DE ET-1. L'ACTION MITOGENE DE ET-1 DISPARAIT EN PRESENCE D'INHIBITEURS SPECIFIQUES DES PKC, APRES LEUR DEGRADATION PAR UN TRAITEMENT PROLONGE PAR LE PHORBOL 12, 13-DIBUTYRATE, ET PAR UNE INHIBITION SELECTIVE DE LA PKC PAR UN OLIGONUCLEOTIDE ANTISENS. DANS LES CELLULES DE LEIOMYOMES, ET-1 POTENTIALISE, VIA L'ACTIVATION DE LA PKC, L'EFFET MITOGENE DE FACTEURS DE CROISSANCE. CETTE ACTION INVERSE DE CELLE OBTENUE DANS LES CELLULES SAINES SUGGERE UN ROLE DE ET-1 DANS LE DEVELOPPEMENT DES TUMEURS BENIGNES DU MYOMETRE. L'ENSEMBLE DE NOS RESULTATS CONTRIBUENT A UNE MEILLEURE CONNAISSANCE DES MECANISMES DE REGULATION PAR ET-1, D'UNE PART DE L'ACTIVITE UTERINE, D'AUTRE PART DU DEVELOPPEMENT NORMAL ET PHYSIOPATHOLOGIQUE DU MYOMETRE HUMAIN. CET APPORT CONSTITUE UN PREALABLE A L'IDENTIFICATION DE CIBLES MOLECULAIRES POTENTIELLES, QUI PERMETTRAIENT DE PROPOSER DE NOUVELLES STRATEGIES THERAPEUTIQUES EFFICACES EN OBSTETRIQUE ET EN GYNECOLOGIE-ONCOLOGIE.PARIS-BIUSJ-Thèses (751052125) / SudocCentre Technique Livre Ens. Sup. (774682301) / SudocPARIS-BIUSJ-Physique recherche (751052113) / SudocSudocFranceF

Gene and protein expression in the myometrium in pregnancy and labor.

“Disclaimer. This is not the definitive version of record of this article. This manuscript has been accepted for publication in Reproduction, but the version presented here has not yet been copy edited, formatted or proofed. Consequently, the journal accepts no responsibility for any errors or omissions it may contain. The definitive version is available at http://dx.doi.org/10.1530/rep.1.00725. © 2006 Society for Reproduction and Fertility.”International audienceMicroarray technologies widen our comprehension of the major structural and metabolic transformations which affect the myometrium from the very beginning of pregnancy until parturition. The results are coherent with the mass of information which was accumulated previously, primarily on the basis of studies of selected critical factors. They highlight the activation of precise signaling pathways, some of which may have been previously under evaluated. The remodelling and maturation processes that the myometrium undergoes in pregnancy appear clearly as phenomena which last during the full course of gestation. Comparatively, the onset of labor is perhaps the phenomenon which remains the least well described by these methods of analysis. Nevertheless, genomic studies constitute a necessary first step of orientation and help establishing new links between the generic signaling pathways that are activated during the normal or pathological gestation. These studies also represent an indicative step that will have to be paralleled, in the future, with the results of the systematic proteomic analysis of the myometrium

Functional genomics of the pregnant uterus: from expectations to reality, a compilation of studies in the myometrium.

ABSTRACT : BACKGROUND : Studies on the human myometrium have reported on different microarrays containing different sets of genes or ESTs. However each study profiled only a small number of patients due to various constraints. More profiling information would be an addition to our knowledge base of parturition. METHODS : We compiled from five human studies, transcriptional differences between the non pregnant myometrium (NP), preterm myometrium (PTNIL), term myometrium not in labor (TNIL) and term myometrium in labor (TIL). Software modules developed by the Draghici's group at Wayne State University (Detroit, MI, USA) were used to propose a hierarchical list of several KEGG pathways most likely adjusted to changes observed in microarray experiments. RESULTS : The differential expression of 118 genes could be dispatched in 14 main KEGG pathways that were the most representative of the changes seen in NP and PTNIL, versus TNIL or TIL. Despite the potential of multiple pitfalls inherent to the use of the microarray technology, gene module analysis of the myometrial transcriptome reveals the activation of precise signaling pathways, some of which may have been under evaluated. CONCLUSION : The remodelling and maturation processes that the uterus undergoes in pregnancy appear clearly as phenomena which last during the full course of gestation. It is attested by the nature of the main signaling pathways represented, in the comparison of the PTNIL versus TNIL uterus. Comparatively, the onset of labor is a phenomenon which remains less well characterized by these methods of analysis, possibly because it is a phenomenon occurring in too short a window to have been grasped by the studies carried out up to now