Table 1.
Primers for qPCR.
Table 2.
Patient demographics for myometrium & placenta biopsies of the RNA-seq cohort—Grouped by phases of labour as determined by cervical dilatation.
Table 3.
Patient demographics for myometrium & placenta biopsies of the RNA-seq cohort—Grouped by status of fetal membrane rupture (ROM).
Table 4.
Patient demographics for myometrium, placenta & choriodecidua biopsies used as independent ‘second cohort’ for qPCR validation of RNA-seq findings.
Fig 1.
PCA for RNA-seq of myometrium biopsies from term pregnant women at different stages of labour.
Summary by principal component analysis (PCA) of overall differences in transcriptome profiles from human myometrium biopsies obtained from term gestation singleton pregnant women, which were grouped by status of cervical (Cx) dilatation or fetal membrane rupture (ROM; >1 hour prior to fetal delivery) at time of Caesarean section; sample identification numbers are shown next to their respective data points. For Cx dilatation, labouring women were grouped as either in early (≤3 cm; TEaL, n = 8) or established (>3 cm; TEsL, n = 6) labour. For ROM, women were grouped as either labouring in the absence (TL-ROM, n = 8) or presence (TL+ROM, n = 6) of ROM. The non-labouring (TNL, n = 8) group was the same for both sets of comparisons.
Table 5.
Number of differentially expressed genes in human myometrium identified by RNA-seq for different labour states.
Table 6.
Top five enriched gene ontology (biological processes) terms for human myometrium in different labour states.
Fig 2.
Grouping human myometrium FC≥1.5 DEGs according to cervical dilatation or ROM status at labour.
Venn diagrams generated from lists of labour-associated up-regulated (↑) and down-regulated (↓) differentially expressed genes (DEGs) in myometrium biopsies, which were obtained from term gestation singleton pregnant women. Values shown only represent number of DEGs that demonstrated median fold change (FC) ≥1.5, when compared to the non-labouring state (TNL, n = 8), in transcript abundance for each of type of labour, which was classified by status of cervical dilatation or fetal membrane rupture (ROM; >1 hour prior to fetal delivery) at time of Caesarean section. For cervical dilatation, labouring women were grouped as either in early (≤3 cm; TEaL, n = 8) or established (>3 cm; TEsL, n = 6) labour. For ROM, women were grouped as either labouring in the absence (TL-ROM, n = 8) or presence (TL+ROM, n = 6) of ROM.
Table 7.
RNA-seq summary for differentially expressed genes common to four labour classifications in human myometrium.
Fig 3.
Relative mRNA abundance determined by qPCR for transcriptomics-identified myometrial DEGs common to four labour classifications in RNA-seq cohort tissues.
Log10 transformed qPCR data (mean with standard deviation; n = 6–8) for myometrial mRNA levels of genes identified by RNA-seq (Fig 2) to be differentially expressed (DEGs) for all four classifications of labour, when compared to the non-labouring state (TNL), in the same biopsies. For cervical dilatation, labour was classified as early (≤3 cm; TEaL) or established (>3 cm; TEsL) at time of Caesarean section. These TEaL and TEsL samples were alternatively classified by fetal membrane rupture (ROM) status, whereby either ROM was absent (TL-ROM) or present (TL+ROM) for >1 hour prior to fetal delivery, to assess the effect of ROM irrespective of cervical dilatation. All data for DEGs of interest were normalised to both β2-microglobulin (B2M) and ribosomal protein L19 (RPL19). Brown-Forsythe & Welch ANOVA (Dunnett’s T3 post-hoc) was used for statistical analyses of TEaL / TEsL vs TNL (a p≤0.05, aa p≤0.01) and TL-ROM / TL+ROM vs TNL (b p≤0.05, bb p≤0.01).
Fig 4.
Relative mRNA abundance determined by qPCR for transcriptomics-identified myometrial DEGs common to TEaL in second cohort tissues.
Log10 transformed qPCR data (mean with standard deviation; n = 7–8) for myometrial mRNA levels of genes identified by RNA-seq (Fig 2) to be differentially expressed (DEGs) during early phase labour (defined as ≤3 cm cervical dilatation; TEaL), when compared to the non-labouring state (TNL), in biopsies obtained from a cohort of term gestation singleton pregnant women separate to those in the RNA-seq cohort. All data for DEGs of interest were normalised to both β2-microglobulin (B2M) and ribosomal protein L19 (RPL19). Two-tailed Welch’s t-test was used for statistical analysis of TEaL vs TNL (** p≤0.01, **** p≤0.0001).
Fig 5.
Placenta and choriodecidua tissue expression levels for transcriptomics-identified myometrial DEGs common to four labour classifications.
Log10 transformed qPCR data (mean with standard deviation) for mRNA expression levels in placenta and choriodecidua biopsies of genes identified from myometrium RNA-seq data (Fig 2) to be differentially expressed (DEGs) for four classifications of labour, when compared to the non-labouring state (TNL), in biopsies from the RNA-seq (‘#1’; n = 5–8) and second (‘#2’; n = 7–8) cohorts of term gestation singleton pregnant women. For cervical dilatation, labour was classified as early (≤3 cm; TEaL) or established (>3 cm; TEsL) at time of Caesarean section. These TEaL and TEsL samples were alternatively classified by fetal membrane rupture (ROM) status, whereby either ROM was absent (TL-ROM) or present (TL+ROM) for >1 hour prior to fetal delivery, to assess the effect of ROM irrespective of cervical dilatation. Data for all DEGs of interest were normalised to both β2-microglobulin (B2M) and ribosomal protein L19 (RPL19). For statistical analyses of cohort #1 data, Brown-Forsythe & Welch ANOVA (Dunnett’s T3 post-hoc) was used for TEaL / TEsL vs TNL and TL-ROM / TL+ROM vs TNL; all p>0.05. For statistical analysis of cohort #2 data, Welch’s t-test was used for TEaL vs TNL; all p>0.05.