H3K27me3 dynamics dictate evolving uterine states in pregnancy and parturition
Patrice Nancy, Johan Siewiera, Gabrielle Rizzuto, Elisa Tagliani, Ivan Osokine, Priyanka Manandhar, Igor Dolgalev, Caterina Clementi, Aristotelis Tsirigos, Adrian Erlebacher
Patrice Nancy, Johan Siewiera, Gabrielle Rizzuto, Elisa Tagliani, Ivan Osokine, Priyanka Manandhar, Igor Dolgalev, Caterina Clementi, Aristotelis Tsirigos, Adrian Erlebacher
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Research Article Reproductive biology

H3K27me3 dynamics dictate evolving uterine states in pregnancy and parturition

Abstract

Uncovering the causes of pregnancy complications such as preterm labor requires greater insight into how the uterus remains in a noncontractile state until term and then surmounts this state to enter labor. Here, we show that dynamic generation and erasure of the repressive histone modification tri-methyl histone H3 lysine 27 (H3K27me3) in decidual stromal cells dictate both elements of pregnancy success in mice. In early gestation, H3K27me3-induced transcriptional silencing of select gene targets ensured uterine quiescence by preventing the decidua from expressing parturition-inducing hormone receptors, manifesting type 1 immunity, and most unexpectedly, generating myofibroblasts and associated wound-healing responses. In late gestation, genome-wide H3K27 demethylation allowed for target gene upregulation, decidual activation, and labor entry. Pharmacological inhibition of H3K27 demethylation in late gestation not only prevented term parturition, but also inhibited delivery while maintaining pup viability in a noninflammatory model of preterm parturition. Immunofluorescence analysis of human specimens suggested that similar regulatory events might occur in the human decidua. Together, these results reveal the centrality of regulated gene silencing in the uterine adaptation to pregnancy and suggest new areas in the study and treatment of pregnancy disorders.

Authors

Patrice Nancy, Johan Siewiera, Gabrielle Rizzuto, Elisa Tagliani, Ivan Osokine, Priyanka Manandhar, Igor Dolgalev, Caterina Clementi, Aristotelis Tsirigos, Adrian Erlebacher

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Figure 4

H3K27 demethylation in late-gestation DSCs, associated with gene upregulation.

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H3K27 demethylation in late-gestation DSCs, associated with gene upregul...
RNA-Seq (A and F) and ChIP-Seq (B and F) were performed on purified cells cultured for 24 hours (n = 3 samples/group). (A) RNA-Seq analysis of various H3K27me3 DSC>MSC targets and inducible inflammatory targets. Other genes of interest are also shown. (B) Distribution of H3K27me3 ±5 kb around the TSS of DSC>MSC targets. Replicates are shown. (C and D) Representative Western blots (C) and normalized H3K27me3:total H3 levels (D) of cells isolated from E15.5 mice injected daily with GSK-J4 or vehicle (–) from E13.5 and cultured for 24 hours (mean ± SEM of n = 4 samples/group; 2 sets of blots). For each set, the average H3K27me3/total H3 ratio for the MSC samples from vehicle-treated mice was set to 1.0 (P = 0.0004, 1-way ANOVA; *P = 0.012; **P < 0.001). Of note, E15.5 DSCs contained less total H3 than MSCs; GSK-J4 had no effect on levels of H3K9me3, another repressive histone mark (Supplemental Figure 6, C–G). (E) H3K27me3 demethylase activity measured on nuclear extracts of whole tissue or tissue layers (mean ±S EM; n = 3 mice/group). The spike in activity in the E15.5 decidua was significantly different from all other groups (P < 0.0001, 1-way ANOVA; *P < 0.05 compared with all other groups). (F) Relationship between H3K27me3 status in DSCs and gene-expression changes (mean ± SEM; n = 3 samples/group). Significantly different DSC groups are indicated (P < 0.0001, 1-way ANOVA; ****P < 0.0001 compared with all other DSC groups). Genes with undetectable H3K27me3 on E7.5 showed no average expression change from E7.5 to E15.5, as did genes for which H3K27me3, while detectable, was either insignificantly changed or reduced less than 1.5 log2–fold. In contrast, greater degrees of H3K27me3 loss correlated with increasingly greater degrees of upregulation. This pattern was not evident for MSCs. Each DSC group was significantly different from its respective MSC group (P < 0.0001, paired t test).