Transcriptomic responses to hypoxia in endometrial and decidual stromal cells

in Reproduction

Correspondence should be addressed to K T Rytkönen; Email: katury@utu.fi
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Human reproductive success depends on a properly decidualized uterine endometrium that allows implantation and the formation of the placenta. At the core of the decidualization process are endometrial stromal fibroblasts (ESF) that differentiate to decidual stromal cells (DSC). As variations in oxygen levels are functionally relevant in endometrium both upon menstruation and during placentation, we assessed the transcriptomic responses to hypoxia in ESF and DSC. In both cell types, hypoxia-upregulated genes in classical hypoxia pathways such as glycolysis and the epithelial mesenchymal transition. In DSC, hypoxia restored an ESF-like transcriptional state for a subset of transcription factors that are known targets of the progesterone receptor, suggesting that hypoxia partially interferes with progesterone signaling. In both cell types, hypoxia modified transcription of several inflammatory transcription factors that are known regulators of decidualization, including decreased transcription of STATs and increased transcription of CEBPs. We observed that hypoxia-upregulated genes in ESF and DSC had a significant overlap with genes previously detected to be upregulated in endometriotic stromal cells. Promoter analysis of the genes in this overlap suggested the hypoxia-upregulated Jun/Fos and CEBP transcription factors as potential drivers of endometriosis-associated transcription. Using immunohistochemistry, we observed increased expression of JUND and CEBPD in endometriosis lesions compared to healthy endometria. Overall, the findings suggest that hypoxic stress establishes distinct transcriptional states in ESF and DSC and that hypoxia influences the expression of genes that contribute to the core gene regulation of endometriotic stromal cells.

Supplementary Materials

    • Supplementary Figure 1. Simplified diagram depicting the decidualization protocol and hypoxia exposures. As hypoxia considerably slows down cellular processes we defined that normoxic decidualization of 40 hours followed by 24 hours decidualization in hypoxia (1% O2) represents a reasonable approximation to be compared to normoxic decidualization of 48 hours. ESF = endometrial stromal fibroblasts, DSC = decidual stromal cells, cAMP = 8-bromoadenosine 3′,5′-cyclic monophosphate (8-Br-cAMP), MPA = medroxyprogesterone acetate.
    • Supplementary Figure 2. The PCA analysis of the four conditions (ESF, DSC, ESF hypoxia and DSC hypoxia) after edgeR upper quartile normalization displays that the two biological replicates inside each experimental condition group tightly together. PC1 separates hypoxia and normoxia conditions and PC2 separates differentiation state (ESF or DSC).
    • Supplementary Figure 3. Subset of Glycolysis pathway showing hypoxia upregulated enzymes in both ESF and DSC (in red, FDR < 0.01, FC > 2.0, TPM > 2).
    • Supplementary Table 1
    • Supplementary Table 2
    • Supplementary Table 3
    • Supplementary Table 4

 

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