Project description:Miscarriage occurs in 15-20% of clinical pregnancies. While chromosomal errors are observed in over 50%, causes of karyotypically normal losses are poorly understood. DNA methylation undergoes reprogramming during development and must be appropriately set to maintain a healthy pregnancy. We hypothesize that aberrant DNA methylation may cause karyotypically normal miscarriage, particularly among women experiencing recurrent miscarriage (RM). DNA methylation in first trimester chorionic villi was assessed in chromosomally normal miscarriages from women with RM (N=33) or isolated miscarriage (M, N=21), and elective terminations (TA, N=16). Differentially methylated candidate loci were identified using the Illumina Infinium HumanMethylation27 BeadChip array by comparing 10 RM to 10 TA samples. Follow up showed a significant increase in methylation in RM and M compared to TA placentae at the CYP1A2 (p=0.002) and AXL (p=0.02) promoters and decrease at the DEFB1 (p=0.008) promoter. Gene function analysis showed an enrichment of imprinted genes (p=9.53E-10) and genes previously associated with RM (p=9.51E-06). DNA methylation was evaluated at 7 imprinted loci using bisulfite pyrosequencing. An increase of outliers at these loci was observed in RM (3.9%) compared to M (0%) and TA (0.9%) (p=0.02), with increased average methylation at the H19/IGF2 ICR1 in M samples (p<0.0001). Altered DNA methylation in the placenta at specific loci, as well as global dysregulation in specific cases, may contribute to or be a consequence of placental insufficiency in karyotypically normal miscarriage. First-trimester placental villi samples from karyotypically normal miscarriages from recurrent miscarriage patients (N, N=10) and chromosomally normal elective terminations (PZET, N=10).
Project description:Whole-transcriptome profiles of individual human placental villi samples from twenty-five (25) Indian women with normal pregnancies during 6- to 8-weeks of gestation were examined using human whole genome expression arrays (NimbleGen 135K). The present study focused on the whole-transcriptome profiling using NimbleGen135K (070925_HG18_exp__12X135K) human whole genome expression arrays of individual human placental villi samples obtained from twenty-five (25) proven-fertile women bearing normal pregnancies voluntarily terminated between 6- and 8-weeks of gestation. Gestational age was estimated from menstrual history, physical and ultrasonographic evaluation. No case of complicated pregnancy from infection, and other significant fetal and maternal clinical indications was included. These twenty five (25) samples include biological replicates of 6, 7 and 8 weeks placental villi samples.
Project description:Whole-transcriptome profiles of individual human placental villi samples from twenty-five (25) Indian women with normal pregnancies during 6- to 8-weeks of gestation were examined using human whole genome expression arrays (NimbleGen 135K).
Project description:Miscarriage occurs in 15-20% of clinical pregnancies. While chromosomal errors are observed in over 50%, causes of karyotypically normal losses are poorly understood. DNA methylation undergoes reprogramming during development and must be appropriately set to maintain a healthy pregnancy. We hypothesize that aberrant DNA methylation may cause karyotypically normal miscarriage, particularly among women experiencing recurrent miscarriage (RM). DNA methylation in first trimester chorionic villi was assessed in chromosomally normal miscarriages from women with RM (N=33) or isolated miscarriage (M, N=21), and elective terminations (TA, N=16). Differentially methylated candidate loci were identified using the Illumina Infinium HumanMethylation27 BeadChip array by comparing 10 RM to 10 TA samples. Follow up showed a significant increase in methylation in RM and M compared to TA placentae at the CYP1A2 (p=0.002) and AXL (p=0.02) promoters and decrease at the DEFB1 (p=0.008) promoter. Gene function analysis showed an enrichment of imprinted genes (p=9.53E-10) and genes previously associated with RM (p=9.51E-06). DNA methylation was evaluated at 7 imprinted loci using bisulfite pyrosequencing. An increase of outliers at these loci was observed in RM (3.9%) compared to M (0%) and TA (0.9%) (p=0.02), with increased average methylation at the H19/IGF2 ICR1 in M samples (p<0.0001). Altered DNA methylation in the placenta at specific loci, as well as global dysregulation in specific cases, may contribute to or be a consequence of placental insufficiency in karyotypically normal miscarriage.
Project description:This study was designed to investigate the miRNA expression profilein human chorionic villi from early recurrent miscarriage. We identified 155 miRNAs exhibiting over 2-fold change in expression levels (P<0.05) by microarray,
Project description:The project aimed to determine proteomics changes in chorionic villi from patients suffering from unexplained early recurrent pregnancy loss (RPL) compared to women undergoing elective abortions. Comparative proteomics analysis by label-free data-independent LC-MS/MS was performed on fresh frozen chorionic villi from 13 RPL patients and 10 controls. Samples from EPL patients were preselected to include only patients with 2 or more recurrent pregnancy losses (RPL), no previous live birth, gestational week from 6-10 weeks and excluded fetal chromosomal abnormalities. Control samples were from women without history of previous miscarriage, ectopic pregnancy or preterm delivery, with at least one live born child, at 6-10 gestational weeks and without fetal chromosomal abnormalities. Patients and controls were between 24-44 years with no significant differences among groups regarding age (Median age RPL = 29; Median age Controls = 34; Mann–Whitney U-test p = 0.078). The mean gestational weak (GW) was 7.9±0.8 weeks in the RPL group and 8.3±1.2 weeks in the control group, without statistically significant difference between groups (Median GW RPL = 8; Median GW Controls = 8; Mann–Whitney U-test p = 0.484).
Project description:Genome wide DNA methylation profiling of chorionic villi and decidua from recurrent miscarriage patients and artificial abortion controls. Infinium HumanMethylation450 BeadChip was used.
Project description:We profiled the transcriptomes of about 110000 single cells from 16 human first-trimester decidual and villi samples from 5 normal samples and 3 recurrent miscarriage (RM) samples. Each decidual and villi samples was collected from the same patient. The cellular composition revealed five major subsets of immune cells including NK, T cell, macrophage, monocytes and B cell in the maternal fetal interface. We used the marker gene of macrophages to establish the diagnosis and treatment model of unexplained recurrent abortion, in order to explore the relationship between immune cells and unexplained recurrent abortion.
Project description:Placental insufficiency is implicated in spontaneous preterm birth (SPTB). We performed RNA-seq study in male and female placentas from women (African American, self-identified) with SPTB (< 36 weeks gestation) compared to normal pregnancies (≥ 38 weeks gestation) to assess the alterations in gene expression profiles.