Bisphenol-A (BPA) is a nonsteroidal estrogen that is ubiquitous in the environment. The homeobox gene Hoxa10 controls uterine organogenesis, and its expression is affected by in utero BPA exposure. We hypothesized that an epigenetic mechanism underlies BPA-mediated alterations in Hoxa10 expression. We analyzed the expression pattern and methylation profile of Hoxa10 after in utero BPA exposure. Pregnant CD-1 mice were treated with BPA (5 mg/kg IP) or vehicle control on d 9-16 of pregnancy. Hoxa10 mRNA and protein expression were increased by 25% in the reproductive tract of mice exposed in utero. Bisulfite sequencing revealed that cytosine-guanine dinucleotide methylation was decreased from 67 to 14% in the promoter and from 71 to 3% in the intron of Hoxa10 after in utero BPA exposure. Decreased DNA methylation led to an increase in binding of ER-alpha to the Hoxa10 ERE both in vitro as and in vivo as determined by EMSA and chromatin immunoprecipitation, respectively. Diminished methylation of the ERE-containing promoter sequence resulted in an increase in ERE-driven gene expression in reporter assays. We identify altered methylation as a novel mechanism of BPA-induced altered developmental programming. Permanent epigenetic alteration of ERE sensitivity to estrogen may be a general mechanism through which endocrine disruptors exert their action.

Diethylstilbestrol (DES) and bisphenol-A (BPA) are estrogen-like endocrine-disrupting chemicals that induce persistent epigenetic changes in the developing uterus. However, DES exposure in utero is also associated with an increased risk of breast cancer in adult women. Similarly, fetal exposure to BPA induces neoplastic changes in mammary tissue of mice. We hypothesized that epigenetic alterations would precede the increased risk of breast neoplasia after in utero exposure to endocrine disruptors. Enhancer of Zeste Homolog 2 (EZH2) is a histone methyltransferase that has been linked to breast cancer risk and epigenetic regulation of tumorigenesis. We examined the effect of BPA and DES on EZH2 expression and function in MCF-7 cells and in mammary glands of mice exposed in utero. DES and BPA treatment approximated human exposure. EZH2 functional activity was assessed by measuring histone H3 trimethylation. Treatment of MCF-7 cells with DES or BPA led to a 3- and 2-fold increase in EZH2 mRNA expression, respectively (p<0.05) as well as increased EZH2 protein expression. Mice exposed to DES in utero showed a >2-fold increase in EZH2 expression in adult mammary tissue compared with controls (p<0.05). EZH2 protein was elevated in mammary tissue of mice exposed to DES or BPA. Histone H3 trimethylation was increased in MCF-7 cells treated with BPA or DES. Similarly, mice exposed to BPA or DES in utero showed increased mammary histone H3 trimethylation. Developmental programming of EZH2 is a novel mechanism by which in utero exposure to endocrine disruptors leads to epigenetic regulation of the mammary gland.

An abnormally decreased placental weight has been linked to increased perinatal complications, including intrauterine fetal demise (IUFD) and fetal growth restriction (IUGR). Despite its promise, determining placental weight prenatally using three-dimensional systems is time-consuming and requires expensive technology and expertise. We propose a novel method using two-dimensional sonography that provides an immediate estimation of placental volume. Placental volume was calculated in 29 third-trimester pregnancies using linear measurements of placental width, height, and thickness to calculate the convex-concave shell volume within 24 hours of birth. Data were analyzed to calculate Spearman's rho (r (s)) and significance. There was a significant correlation between estimated placental volume (EPV) and actual placental weight (r (s) = 0.80, P < 0.001). Subgroup analysis of preterm gestations ( N = 14) revealed an even more significant correlation of EPV to actual placental weight (r (s) = 0.89, P < 0.001). Placental weight can be accurately predicted by two-dimensional ultrasound with volumetric calculations. This method is simple, rapid, and accurate, making it practical for routine prenatal care, as well as for high-risk cases with decreased fetal movement and IUGR. Routine EPV surveillance may decrease the rates of perinatal complications and unexpected IUFD.

Objective To determine if TGF-β3 is a paracrine signal secreted by leiomyoma that inhibits BMP mediated endometrial receptivity and decidualization. Design Experimental Setting Laboratory Patients Women with symptomatic leiomyomas Interventions Endometrial stromal cells (ESC) and leiomyoma cells were isolated from surgical specimens. Leiomyoma-conditioned media (LCM) was applied to cultured ESC. TGF-β was blocked by two approaches: TGF-β pan-specific antibody or transfection with a mutant TGF-β receptor type II. Cells were then treated with rhBMP-2 to assess BMP responsiveness. Main Outcome Measures Expression of BMP receptor types 1A (BMPR1A), 1B (BMPR1B), 2 (BMPR2), as well as endometrial receptivity mediators HOXA10 and LIF. Results ELISA showed elevated TGF-β levels in LCM. LCM treatment of ESC reduced expression of BMPR1B and BMPR2 to approximately 60% of pretreatment levels. Pre-incubation of LCM with TGF-β neutralizing antibody or mutant TGF receptor, but not respective controls, prevented repression of BMP receptors. HOXA10 and LIF expression was repressed in rhBMP-2 treated, LCM exposed ESC. Pre-treatment of LCM with TGF-β antibody or transfection with mutant TGF receptor prevented HOXA10 and LIF repression. Conclusions Leiomyoma derived TGF-β was necessary and sufficient to alter endometrial BMP-2 responsiveness. Blockade of TGF-β prevents repression of BMP-2 receptors and restores BMP-2 stimulated expression of HOXA10 and LIF. Blockade of TGF signaling is a potential strategy to improve infertility and pregnancy loss associated with uterine leiomyoma.

Prolonged (postterm) pregnancies, although less common in the era of ultrasound dating, are associated with fetal and maternal risks. The authors favor routine induction of labor at 41 weeks in women with low-risk singleton gestations.