Vitamin D Regulates Maternal T-Helper Cytokine Production in Infertile Women

Vitamin D (VD) deficiency is associated with reproductive failure. However, the relationship between VD and maternal immunity remains unclear. We investigated the clinical efficacy of VD in maternal T-helper (Th) cytokines in 276 infertile women and examined for Th1 and Th2 cells based on the deficient, insufficient, and sufficient serum 25-hydroxyvitamin D3 (25[OH]VD) levels (<12, 12–30, and >30 ng/mL, respectively). Most infertile women had a low-level of VD (87.3%). Immunological tests of pre-/post-VD supplementation were performed in patients who were deficient and insufficient in VD. Of 23 patients, 11 (47.8%) exhibited sufficient VD levels after supplementation. Th1/Th2 cell ratio in patients with insufficient VD was significantly decreased after supplementation (p = 0.004). After supplementation, serum 25(OH)VD levels of the patients: 11 in the sufficient group showed significant decreases in Th1 cell level and Th1/Th2 cell ratio (p = 0.032 and 0.010, respectively), whereas no significant differences in Th1/Th2 cell ratio were recognized in the insufficient group. Furthermore, mid-luteal endometrial biopsies (n = 18) were processed for primary cultures and measured interferon [IFN]-γ and interleukin [IL]-4 in condition media. Decidualizing cultures with 1,25-dihydroxvitamin D3 (1,25[OH]2VD) decreased IFN-γ. Sufficient VD supplementation in women with insufficient VD may optimize maternal T-helper cytokines during pregnancy via rebalancing the Th1/Th2 cell ratio

Resveratrol inhibits decidualization by accelerating downregulation of the CRABP2-RAR pathway in differentiating human endometrial stromal cells

Abstract Pregnancy critically depends on the transformation of the human endometrium into a decidual matrix that controls embryo implantation and placenta formation, a process driven foremost by differentiation and polarization of endometrial stromal cells into mature and senescent decidual cells. Perturbations in the decidual process underpin a spectrum of prevalent reproductive disorders, including implantation failure and early pregnancy loss, emphasizing the need for new therapeutic interventions. Resveratrol is a naturally occurring polyphenol, widely used for its antioxidant and anti-inflammatory properties. Using primary human endometrial stromal cell (HESC) cultures, we demonstrate that resveratrol has anti-deciduogenic properties, repressing not only the induction of the decidual marker genes PRL and IGFBP1 but also abrogating decidual senescence. Knockdown of Sirtuin 1, a histone deacetylase activated by resveratrol, restored the expression of IGFBP1 but not the induction of PRL or senescence markers in decidualizing HESCs, suggesting involvement of other pathways. We demonstrate that resveratrol interferes with the reprogramming of the retinoic acid signaling pathway in decidualizing HESCs by accelerating down-regulation of cellular retinoic acid-binding protein 2 (CRABP2) and retinoic acid receptor (RAR). Notably, knockdown of CRABP2 or RAR in HESCs was sufficient to recapitulate the anti-deciduogenic effects of resveratrol. Thus, while resveratrol has been advanced as a potential fertility drug, our results indicate it may have detrimental effects on embryo implantation by interfering with decidual remodeling of the endometrium

Tables for fpkm values

Tables for fpkm values for RNA-seq data calculated using Cufflinks:genes.fpkm_tracking, isoforms.fpkm_tracking, and tss_groups.fpkm_trackin

Peak call files for ChIP-seq data

Peak call files for CHIP-seq data produced by MACS2:EM0409D0_K27ac_macs2_peaks.narrowPeak EM0409D0_K27me3_macs2_peaks.broadPeak EM0409D0_K9me3_macs2_peaks.broadPeak EM0409D4_K27ac_macs2_peaks.narrowPeak EM0409D4_K27me3_macs2_peaks.broadPeak EM0409D4_K9me3_macs2_peaks.broadPeak EM0409D8_K27ac_macs2_peaks.narrowPeak EM0409D8_K27me3_macs2_peaks.broadPeak EM0409D8_K9me3_macs2_peaks.broadPeak EM0519D0_K27ac_macs2_peaks.narrowPeak EM0519D0_K27me3_macs2_peaks.broadPeak EM0519D0_K9me3_macs2_peaks.broadPeak EM0519D4_K27ac_macs2_peaks.narrowPeak EM0519D4_K27me3_macs2_peaks.broadPeak EM0519D4_K9me3_macs2_peaks.broadPeak EM0519D8_K27ac_macs2_peaks.narrowPeak EM0519D8_K27me3_macs2_peaks.broadPeak EM0519D8_K9me3_macs2_peaks.broadPea

Data from: Reciprocal changes of H3K27ac and H3K27me3 at the promoter regions of the critical genes for endometrial decidualization

Aim: Decidualization is essential for embryo implantation and placental development. We aimed to obtain transcriptome and epigenome profiles for primary endometrial stromal cells (ESCs) and in vitro decidualized cells. Materials & methods: ESCs isolated from human endometrial tissues remained untreated (D0), or decidualized for 4 days (D4) and 8 days (D8) in the presence of 8-bromo-cAMP and progesterone. Results: Among the epigenetic modifications examined (DNA methylation, H3K27ac, H3K9me3 and H3K27me3), the H3K27ac patterns changed most dramatically, with a moderate correlation with gene expression changes, upon decidualization. Subsets of up- and down-regulated genes upon decidualization were associated with reciprocal changes of H3K27ac and H3K27me3 modifications at their promoter region, and were enriched with genes essential for decidualization such as WNT4, ZBTB16, PROK1 and GREB1. Conclusion: Our dataset is useful to further elucidate the molecular mechanisms underlying decidualization

bigwig_files

"bigwig_files.zip" (md5 checksum: a7cee3f55c6c3d53dd562475aa7fc1f3). The "bigwig_files.zip" file contains the following 25 bigwig files (19 files for ChIP-seq data and 6 files for RNA-sea data). EM0409_D0_H3K27ac.hg19.bw EM0409_D0_H3K27me3.hg19.bw EM0409_D0_H3K9me3.hg19.bw EM0409_D4_H3K27ac.hg19.bw EM0409_D4_H3K27me3.hg19.bw EM0409_D4_H3K9me3.hg19.bw EM0409_D4_input.hg19.bw EM0409_D8_H3K27ac.hg19.bw EM0409_D8_H3K27me3.hg19.bw EM0409_D8_H3K9me3.hg19.bw EM0519_D0_H3K27ac.hg19.bw EM0519_D0_H3K27me3.hg19.bw EM0519_D0_H3K9me3.hg19.bw EM0519_D4_H3K27ac.hg19.bw EM0519_D4_H3K27me3.hg19.bw EM0519_D4_H3K9me3.hg19.bw EM0519_D4_input.hg19.bw EM0519_D8_H3K27me3.hg19.bw EM0519_D8_H3K9me3.hg19.bw RNAseq_EM0409_D0.hg19.bw RNAseq_EM0409_D4.hg19.bw RNAseq_EM0409_D8.hg19.bw RNAseq_EM0519_D0.hg19.bw RNAseq_EM0519_D4.hg19.bw RNAseq_EM0519_D8.hg19.b

Establishment of a PCR analysis method for canine BRCA2

Abstract Background Mammary tumors are the most common tumor type in both human and canine females. In women, carriers of mutations in BRCA2, a tumor suppressor gene product, have a higher risk of breast cancer. Canine BRCA2 has also been suggested to have a relationship with mammary tumors. However, clearly deleterious BRCA2 mutations have not been identified in any canine mammary tumors, as appropriate methods to detect mutations or a consensus BRCA2 sequence have not been reported. Findings For amplification and sequencing of BRCA2, we designed 14 and 20 PCR primer sets corresponding to the BRCA2 open reading frame (ORF) and all 27 exons, respectively, including exon-intron boundaries of the canine BRCA2 regions, respectively. To define the consensus canine BRCA2 ORF sequence, we used established methods to sequence the full-length canine BRCA2 ORF sequence from two ovaries and a testis obtained from individual healthy mongrel dogs and partially sequence BRCA2 genomic sequences in 20-56 tumor-free dogs, each aged over 6 years. Subsequently, we compared these sequences and seven previously reported sequences, and defined the most common base sequences as the consensus canine BRCA2 ORF sequence. Moreover, we established a detection method for identifying splicing variants. Unexpectedly, we also identified novel splicing variants in normal testes during establishment of these methods. Conclusions The present analysis methods for determining the BRCA2 base sequence and for detecting BRCA2 splicing variants and the BRCA2 ORF consensus sequence are useful for better understanding the relationship between canine BRCA2 mutation status and cancer risk.</p

Reprogramming of the retinoic acid pathway in decidualizing human endometrial stromal cells

Upon breaching of the endometrial surface epithelium, the implanting embryo embeds in the decidualizing stroma. Retinoic acid (RA), a metabolite of vitamin A, is an important morphogen during embryonic and fetal development, although the role of the RA pathway in the surrounding decidual cells is not understood. Here we show that decidual transformation of human endometrial stromal cells (HESCs) results in profound reprogramming of the RA signaling and metabolism pathways. Differentiating HESCs downregulate the intracellular carrier proteins CRABP2 and FABP5, responsible for transfer and binding of RA to the nuclear receptors RAR and PPARβ/δ, respectively. Furthermore, the expression of RAR, the receptor that mediates the pro-apoptotic effects of RA, was also inhibited. By contrast, PPARβ/δ, which transduces the differentiation responses of RA, was upregulated. Decidualization was also associated with increased expression of retinol-binding protein 4 (RBP4) and various enzymes involved in the metabolism of RA and its precursor, retinaldehyde (Rald), including CYP26A1, DHRS3, and RDH12. Exposure of differentiating HESCs to RA or Rald reversed the inhibition of the CRABP2-RAR pathway, perturbed the expression of decidual marker genes and triggered cell death. Taken together, the data demonstrate that decidualizing HESCs silence RA signaling by downregulating key cytoplasmic binding proteins and by increasing retinoid metabolism. However, excessive RA exposure is toxic for decidual cells and triggers a response that may lead to pregnancy failure