Autism-Like Behavior in the Offspring of CYP11A1-Overexpressing Pregnant Rats.

Autism spectrum disorders (ASD) represent a complex group of neurodevelopmental disorders that are characterized by impaired social behavior and communication as well as repetitive behavior and restricted interests. Prenatal exposure to high levels of testosterone and preeclampsia are thought to be risk factors of ASD. We had previously reported that overexpression of the mitochondrial cholesterol side-chain cleavage enzyme (CYP11A1) could lead to both preeclampsia-like symptoms and increased testosterone levels in pregnant rats. In this study, we investigated the association between high CYP11A1 levels in pregnant rats and autism-like behavior in their offspring. Timed-pregnant Sprague-Dawley (SD) rats were injected with CYP11A1 gene-carrying adenoviruses on gestational day 8.5, and their offspring were then compared with those from timed-pregnant control SD rats. Compared with their control counterparts, the offspring of the CYP11A1-ovexpressing dams displayed more symptoms of anxiety and spent less time in social interactions and more time in self-grooming and rearing, all indicators of autism-like behavior. Sequencing of the transcriptome in primary microglia from the offspring of CYP11A1-overexpressing dams revealed that immune pathways were highly activated, and the gamma-aminobutyric acid type A (GABAA) receptor genes were among the top differentially expressed genes. Using primary microglia cultures generated from neonatal rats, tumor necrosis factor-alpha expression was found to be elevated in the cells transfected with CYP11A1-carrying adenoviruses. Additionally, the offspring of CYP11A1-overexpressing dams displayed dysregulated GABAA receptor expression. Taken together, these results suggest that abnormal CYP11A1 gene expression in pregnant rats could lead to microglial immune activation and dysregulated GABAA receptor expression in their offspring and thereby anxiety and autism-related behavior. Our study suggests that the pathways regulated by CYP11A1 could be promising preventative and therapeutic targets for ASD.

Hepatitis B surface antigen positivity during pregnancy and risk of gestational diabetes mellitus: A systematic review and meta-analysis.

Chronic hepatitis B virus (HBV) infection is a prevalent public health issue worldwide. Its impact on important pregnancy outcomes, such as gestational diabetes mellitus (GDM), has not been clearly established. The findings from published studies are inconsistent. In this systematic review and meta-analysis, we aimed to clarify whether HBV infection manifested during pregnancy is associated with an increased risk of GDM. We searched MEDLINE and EMBASE for cohort studies and case-control studies that investigated the association between maternal hepatitis B surface antigen (HBsAg) positivity and GDM. We pooled adjusted odds ratio (aOR) and unadjusted OR, respectively, using the random-effect generic inverse variance method. We assessed risk of bias using the Quality in Prognosis Studies tool and conducted five pre-specified subgroup analyses. In total, 23 cohort studies involving 3 529 223 participants were included. The risk of GDM was 6.48% (1868/28 829) among HBsAg-positive pregnant women and 3.41% (119 283/3 500 394) among HBsAg-negative pregnant women. Meta-analyses of both unadjusted and adjusted effect estimates showed that HBsAg positivity during pregnancy was associated with higher risk of developing GDM (unadjusted OR 1.35, 95% CI: 1.17 to 1.56, I2  = 82.6%; adjusted OR 1.47, 1.22 to 1.76, I2  = 62%). Among pre-specified subgroup analysis, significant differences were found among studies with high vs low or moderate risk of bias. The results were robust to sensitivity analyses. In conclusion, HBsAg positivity during pregnancy has a moderate effect on an increased risk of GDM. Given the size of the population with HBV infection worldwide, however, this effect could have substantial impact on pregnancy. Further studies are warranted to investigate whether active infection with HBeAg positivity would further elevate the risk of adverse events during pregnancy.

Ten-eleven translocation 2 demethylates the MMP9 promoter, and its down-regulation in preeclampsia impairs trophoblast migration and invasion.

Preeclampsia is the most common clinical disorder in pregnancy and might result from disordered uterine environments caused by epigenetic modifications, including deregulation of DNA methylation/demethylation. Recent research has indicated that 5-hydroxymethylcytosine (5hmC), a DNA base derived from 5-methylcytosine (5mC) via oxidation by ten-eleven translocation (TET) enzymes, is involved in DNA methylation-related plasticity. Here, we report that TET2 expression and 5hmC abundance are significantly altered in the placentas from preeclampsia patients. shRNA-mediated TET2 knockdown (shTET2) reduced trophoblast migration and invasion when cultured in Matrigel. Both real-time PCR of matrix metalloproteinase (MMP)-related transcripts and a human angiogenesis antibody array indicated that TET2 knockdown in trophoblasts inhibits the expression of MMP transcript, of which MMP9 represented one of the most significant TET2 downstream targets. Using an established shTET2 HTR-8/SVneo cell model, we further confirmed alterations of 5hmC levels and MMP9 expression at both mRNA and protein levels. In particular, we found that TET2 bound to and removed 5mC modifications at the MMP9 promoter region. Interestingly, in TET2 knockdown cells, both MMP9 expression and the compromised trophoblast phenotype could be rescued by vitamin C, an activator of TET enzyme activity. Finally, TET2 expression correlated with MMP9 levels in placenta samples from the preeclampsia patients, indicating that TET2 deregulation is critically involved in the pathogenesis of preeclampsia through down-regulation of MMP9 expression. Our findings highlight a critical role of TET2 in regulating trophoblast cell migration through demethylation at the MMP9 promoter, and suggest that down-regulation of the TET2-MMP9-mediated pathway contributes to preeclampsia pathogenesis.

Immunological cytokine profiling identifies TNF-α as a key molecule dysregulated in autistic children.

Recent studies have suggested that the etiology of autism spectrum disorder (ASD) may be caused by immunological factors, particularly abnormalities in the innate immune system. However, it is still unclear which specific cytokines may be of most importance. The current study therefore investigated which cytokines showed altered concentrations in blood in ASD compared with healthy control children and which were also correlated with symptom severity. Our study sample included 32 children diagnosed with ASD and 28 age and sex-matched typically developing children. Autism symptoms were measured using the Autistic Behavior Checklist (ABC) and blood samples were taken from all subjects. We used Milliplex cytokine kits to determine serum concentrations of 11 Th1, Th2 and Th17 related cytokines. Additionally, expression of THRIL (TNFα and hnRNPL related immunoregulatory LincRNA), a long non-coding RNA involved in the regulation of tumor necrosis factor- α (TNF-α), was determined using real-time PCR. Of the 11 cytokines measured only concentrations of TNF-α (p=0.002), IL-1ß (p=0.02) and IL-17a (p=0.049) were significantly increased in ASD children compared to typically developing controls, but only TNF-α concentrations were positively correlated with severity of ASD symptoms on all 5 different ABC sub-scales and were predictive of an ASD phenotype (area under the curve = 0.74). Furthermore, THRIL RNA expression was significantly decreased in ASD children. Our results provide further support for altered innate immunity being an important autism pathogenic factor, with autistic children showing increased blood TNF-α concentrations associated with symptom severity, and decreased expression of the THRIL gene involved in regulating TNF-α.

Abnormal CYP11A1 gene expression induces excessive autophagy, contributing to the pathogenesis of preeclampsia.

OBJECTIVE: In this study, we investigated the exact mechanism by which excessive CYP11A1 expression impairs the placentation process and whether this causes preeclampsia (PE) in an in vivo model. SETTING AND DESIGN: In order to study CYP11A1 overexpression, BeWo cells were transfected with CYP11A1. Pregnenolone, progesterone, and testosterone levels were measured by enzyme linked immunosorbent assays, and levels of autophagy markers were quantified by western blotting and immunofluorescence. Trophoblastic cell invasion was assessed using transwell assays; BeWo cells were treated with testosterone and an androgen receptor (AR) inhibitor (flutamide) to elucidate the invasion mechanism. An adenovirus overexpression rat model was established to investigate CYP11A1 overexpression in vivo and the phenotype was examined. Furthermore, human placenta samples (n = 24) were used to determine whether PE patient placentas showed altered CYP11A1 and autophagy marker expression. RESULTS: BeWo cells overexpressing CYP11A1 had significantly increased levels of pregnenolone, progesterone, and testosterone. Additionally, the expression levels of autophagy markers in CYP11A1-overexpressing BeWo cells were significantly increased. Trophoblast invasion was significantly reduced in CYP11A1-overexpressing cells as well as in cells treated with high testosterone. This reduction could be significantly rescued when cells were pretreated with flutamide. Overexpression of CYP11A1 in rat pregnancies led to PE-like symptoms and an over-activation of the AR-mediated pathway in the placenta. Elevated expression of CYP11A1 and autophagy markers could also be detected in PE placenta samples. CONCLUSIONS: These results suggest that abnormally high expression of CYP11A1 induces trophoblast autophagy and inhibits trophoblastic invasion, which is associated with the etiology of PE.

Breastfeeding initiation, duration and exclusivity in mothers with epilepsy from South West China.

PURPOSES: 1) To study the breastfeeding initiation, duration and exclusivity rates, and common reasons for early weaning in Chinese mothers with epilepsy (MWE); 2) To identify potential perinatal breastfeeding correlations with selected sociodemographic and clinical factors. METHODS: A semi-structured questionnaire was administered to 281 MWE attending hospitals in South-west China from February 2014 to July 2015. Data about breastfeeding behaviors, sociodemographic, obstetric, and epileptic variables were collected. Descriptive analyses, followed by univariate and multivariate logistic regression analyses, were utilized to examine associations with breastfeeding, its duration and exclusivity. RESULTS: Breastfeeding initiation rate in MWE was 59.4%. At 3 months post partum total breastfeeding rate was 49.5% and exclusive breastfeeding rate was 36.3%. At 6 months, about one third (33.1%) of MWE had continued breastfeeding their babies and 12.8% of enrolled infants were exclusively breastfed. During lactation, fear of exposure of the babies to antiepileptic drugs (AEDs) via breast milk, frequent seizures, and insufficient breast milk supply were the commonest reasons for early cessation of breastfeeding. Mothers with epilepsy who had babies delivered at full term were more inclined to breastfeed their babies. Mothers who had gestational non-active epilepsy were more likely to engage in long-term breastfeeding. AED polytherapy was associated with poor breastfeeding behaviors in all aspects. CONCLUSION: MWE in our study had a lower prevalence of breastfeeding than what would be expected in the general population, where approximately 95% breastfeed. Good seizure control and optimal antiepileptic therapy during gestation and lactation were associated with a higher rate of breastfeeding. Targeted intervention programs enhancing antenatal care services and breastfeeding consultation are needed.

miR-15b-AGO2 play a critical role in HTR8/SVneo invasion and in a model of angiogenesis defects related to inflammation.

INTRODUCTION: microRNAs (miRs) have been shown to play critical roles in the regulation of trophoblast and endothelial cell functions, and one significant finding concerning the miR-15/16 family is that most members of this family are highly expressed in endothelial cells and contribute to functions, such as tube formation. The interaction between trophoblast and endothelial cell play an important role in normal placentation process. Therefore, the aims of this study were to investigate the expression of miR-15b in human placenta and to uncover the potential role of miR-15b as well as its target functional loop in trophoblast and endothelial cells. Whether inflammation could modulate the expression of miR-15b and its down-stream target was further investigated. Additionally, the potential link between miR-15b deregulation and preeclampsia was also explored in the placenta of patients diagnosed with preeclampsia. METHODS: The expression of miR-15b was studied in the placental tissue of a normal pregnancy using in situ hybridization, and the effects of miR-15b on proliferation, invasion, and angiogenesis were further explored in vitro using HTR-8/SVneo and HUVEC cell line models. A Lipopolysaccharides (LPS) treatment model in HTR-8/SVneo cell was utilized to explore the mechanism of how LPS treatment could lead to the activation of miR-15b expression. Western blot was used to detect the expression of proteins related to miR-15b mediated pathway in preeclamptic placentas. RESULTS: miR-15b inhibits trophoblast cell invasion and endothelial cell tube formation by suppressing the expression of Argonaute 2 (AGO2), a major miRNA effecter protein. AGO2 is specifically localized to human placenta cytotrophoblast and endothelial cells, and it plays important roles in trophoblast cell invasion and endothelial cell tube formation. LPS treatment may lead to the overexpression of miR-15b and down-regulation of AGO2, which may be involved in shallow trophoblast cell invasion associated with the pathogenesis of preeclampsia. Chromatin immunoprecipitation assay indicates that increased occupancy of AGO2 to miR-15b promoter is responsible for the increased expression of miR-15b under the condition of LPS treatment. Furthermore, preeclamptic placentas have decreased expression of AGO2, but increased expression of miR-15b and TLR-4 compared to normal controls. DISCUSSION: This is the first report about the function of AGO2 in human trophoblast and endothelial cells in the placenta. The data indicates that the aberrant expression of miR-15b contributes to abnormal placentation by targeting AGO2 mRNA. This study provides insight into the potential role of the miR-15b and AGO2 functional loop in the placentation process.