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Gestational diabetes mellitus placentas exhibit epimutations at placental development genes
No full textGestational diabetes mellitus (GDM) is a maternal metabolic disorder that perturbs placental development and increases the risk of offspring short- and long-term metabolic disorders. The mechanisms by which GDM impairs placental development remain poorly understood. Here, we defined the DNA methylome of GDM placentas and determined whether GDM perturbs methylation at genes important for placental development. We conducted an epigenome-wide association study of 42 placentas from pregnancies in the South African Soweto First 1000 days cohort (S1000). Using genome-wide bisulfite sequencing, we compared non-GDM placentas to GDM placentas with similar proportions from obese and non-obese mothers. Compared to non-GDM, GDM placentas exhibited a distinct methylation profile consisting of 12,210 differentially methylated CpGs (DMCs) that mapped to 3,875 genes. Epigenetically altered genes were enriched in Wnt and cadherin signalling pathways, both critical in placentation and embryogenesis. We also defined regional DNA methylation perturbation in GDM placentas at 11 placental development genes. These findings reveal extensive changes to the placental epigenome of GDM pregnancies and highlight perturbation enriched at important placental development genes. These molecular changes represent potential mechanisms for GDM-induced placental effects that may serve as candidate biomarkers for placental, maternal, and foetal health. Using a study design that used similar proportions of obese and non-obese mothers in our case and control pregnancies, we minimized the detection of changes due to obesity alone. Further work will be necessary to investigate the extent of the influence of obesity on these GDM-related placental epigenetic changes.</span
Potential Mechanisms of Developmental Vitamin D Deficiency (DVD)-Induced Metabolic Disruption in Adult CC001xCC011 Male Mice
Get PDFDevelopmental vitamin D deficiency (DVD), or vitamin D deficiency during fetal and early postnatal development, has recently been identified as a significant public health concern. DVD poses a significant threat to offspring metabolic health and is a hypothesized risk factor for adult metabolic syndrome, obesity, and type 2 diabetes. This study aims to characterize the potential mechanisms of DVD-induced metabolic disruption, including disruption of adipose tissue morphology, alterations in energy balance, and expression of growth-regulator genes. This study evaluates the hypothesis that DVD exposure will reduce Grb10 gene expression in PWAT and brain of CC001xCC011 mice and that the reduced Grb10 expression will be positively correlated with elevated adult adiposity and reduced energy expenditure.Bachelor of Science in Public Healt
Genetic determination of vitamin D status in Collaborative Cross mice
Get PDFVitamin D is an essential micronutrient that plays many important roles in the body. Deficiency of vitamin D has been associated with various diseases, including rickets in children, osteomalacia in adults, type 1 and type 2 diabetes. Vitamin D status is affected by both environmental and genetic factors. However, the mechanisms of genetic determination are unclear. In this project, we aim to characterize the genetic determinants of vitamin D status using Collaborative Cross (CC) inbred mice. Genetic differences at 14 genes important for vitamin D metabolism were examined across 16 CC mice. This was achieved by analyzing single nucleotide polymorphisms (SNPs) at the 14 genes using principal component analysis (PCA). Levels of 4 vitamin D metabolites (25(OH)D3, 1,25(OH)2D3, 24,25(OH)2D3 and 1,24,25(OH)3D3) in 4 CC strains (CC001, CC006, CC011 and CC026) were measured and compared across strains. The genetic differences determined from PCA analyses were then associated to basal levels of vitamin D metabolites as well as the response dietary depletion. Basal levels of 25(OH)D3 and 24,25(OH)2D3 were significantly (adj.p<0.5) different among the 4 CC strains. Genes Cyp27b1, Vdr, Ets1, Trim35 were significantly (adj.p<0.5) associated with basal level of 24,25(OH)2D3. In conclusions, this project showed that CC mice are a valuable model to study genetic determination of vitamin D status, due the their genetic diversity and wide range of vitamin D status. We also demonstrated that genetic differences among CC strains determined from PCA analyses can be used to predict their basal level of 24,25(OH)2D3.Bachelor of Science in Public Healt
Potential Mechanisms of Developmental Vitamin D Deficiency (DVD)-Induced Metabolic Disruption in Adult CC001xCC011 Male Mice
Get PDFDevelopmental vitamin D deficiency (DVD), or vitamin D deficiency during fetal and early postnatal development, has recently been identified as a significant public health concern. DVD poses a significant threat to offspring metabolic health and is a hypothesized risk factor for adult metabolic syndrome, obesity, and type 2 diabetes. This study aims to characterize the potential mechanisms of DVD-induced metabolic disruption, including disruption of adipose tissue morphology, alterations in energy balance, and expression of growth-regulator genes. This study evaluates the hypothesis that DVD exposure will reduce Grb10 gene expression in PWAT and brain of CC001xCC011 mice and that the reduced Grb10 expression will be positively correlated with elevated adult adiposity and reduced energy expenditure.Bachelor of Science in Public Healt
USING ENDOMETRIAL ORGANOIDS TO CHARACTERIZE PERSISTENT DES-INDUCED UTERLINE EPITHELIAL CHANGES
Get PDFPrenatal exposure of diethylstilbestrol (DES), a synthetic estrogen, caused female reproductive abnormalities that lead to infertility and increased cancer risks. In mice, neonatal DES exposure induces similar reproductive dysfunction seen in human prenatal exposure. Neonatal mouse DES exposure causes aberrant gene expression and estrogen dependent alterations to the uterine epigenome and persist into adulthood. In particular, these changes result in altered epithelial cell differentiation which develops into adenocarcinoma later in life. The uterine glandular and luminal epithelium goes through cyclic regeneration and is replenished from a population of resident adult stem cells. Thus, we hypothesized that early DES exposure alters the endometrial stem cell and progenitor populations which causes aberrant epithelial differentiation. To study the persistent effects of DES, CD-1 mice were exposed to a daily dose of DES at 1mg/kg from post-natal day (PND) 1-5 and uteri were collected prior to puberty on postnatal day 16 (PND16). An epithelial enriched cell population was isolated for RNA-seq analysis or cultured into endometrial organoids – designed for optimal stem and progenitor cell survival and expansion. Using bulk RNA-seq, we found that at PND16, the epithelial enriched cell population had characteristic DES induced phenotype in tissue (Ltf and Six1 upregulation). Endometrial organoids were able to capture similar DES effects from the in vivo exposure such as aberrant Wnt signaling pathway and basal cell marker expression. To further understand how epithelial cells are altered, single cell RNA and ATAC sequencing was done using both PND16 whole uterine tissue and PND16-derived endometrial organoids. DES-exposed uteri samples had completely different transcriptional landscape compared to the control. The epithelial population of CON and DES exhibited different characteristics. We found that the endometrial organoid culture was entirely epithelial. Subpopulations included stem cell/progenitor, glandular/secretory, and basal cells in both control and DES. There were no stem cell population and fewer progenitor populations identified in DES-exposed organoids. These early findings suggest that neonatal DES exposure alters endometrial progenitor renewal capacity and epithelial differentiation.Master of Scienc
Supplemental Materials for "Hutchins EK, Niu C, Xue J, et al. (2025) Interindividual genetic differences drive discordance between serum calcidiol and calcitriol concentrations in females. Endocrinology (2025). doi: 10.1210/endocr/bqaf138
Get PDFThis work contains supplemental materials for "Hutchins EK, Niu C, Xue J, et al. (2025) Interindividual genetic differences drive discordance between serum calcidiol and calcitriol concentrations in females. Endocrinology (2025). doi: 10.1210/endocr/bqaf138
Supplemental Table 1
Supplemental Figures 1-6
Please cite the manuscript for all use of this work. doi: 10.1210/endocr/bqaf138
Humanized H19/Igf2 locus reveals diverged imprinting mechanism between mouse and human and reflects Silver-Russell syndrome phenotypes
Get PDFGenomic imprinting affects a subset of genes in mammals, such that they are expressed in a monoallelic, parent-of-origin-specific manner. These genes are regulated by imprinting control regions (ICRs), cis-regulatory elements that exhibit allele-specific differential DNA methylation. Although genomic imprinting is conserved in mammals, ICRs are genetically divergent across species. This raises the fundamental question of whether the ICR plays a species-specific role in regulating imprinting at a given locus. We addressed this question at the H19/insulin-like growth factor 2 (Igf2) imprinted locus, the misregulation of which is associated with the human imprinting disorders Beckwith-Wiedemann syndrome (BWS) and Silver-Russell syndrome (SRS). We generated a knock-in mouse in which the endogenous H19/Igf2 ICR (mIC1) is replaced by the orthologous human ICR (hIC1) sequence, designated H19(hIC1) We show that hIC1 can functionally replace mIC1 on the maternal allele. In contrast, paternally transmitted hIC1 leads to growth restriction, abnormal hIC1 methylation, and loss of H19 and Igf2 imprinted expression. Imprint establishment at hIC1 is impaired in the male germ line, which is associated with an abnormal composition of histone posttranslational modifications compared with mIC1. Overall, this study reveals evolutionarily divergent paternal imprinting at IC1 between mice and humans. The conserved maternal imprinting mechanism and function at IC1 demonstrates the possibility of modeling maternal transmission of hIC1 mutations associated with BWS in mice. In addition, we propose that further analyses in the paternal knock-in H19(+/hIC1) mice will elucidate the molecular mechanisms that may underlie SRS.Genomic imprinting affects a subset of genes in mammals, such that they are expressed in a monoallelic, parent-of-origin-specific manner. These genes are regulated by imprinting control regions (ICRs), cis-regulatory elements that exhibit allele-specific differential DNA methylation. Although genomic imprinting is conserved in mammals, ICRs are genetically divergent across species. This raises the fundamental question of whether the ICR plays a species-specific role in regulating imprinting at a given locus. We addressed this question at the H19/insulin-like growth factor 2 (Igf2) imprinted locus, the misregulation of which is associated with the human imprinting disorders Beckwith-Wiedemann syndrome (BWS) and Silver-Russell syndrome (SRS). We generated a knock-in mouse in which the endogenous H19/Igf2 ICR (mIC1) is replaced by the orthologous human ICR (hIC1) sequence, designated H19(hIC1). We show that hIC1 can functionally replace mIC1 on the maternal allele. In contrast, paternally transmitted hIC1 leads to growth restriction, abnormal hIC1 methylation, and loss of H19 and Igf2 imprinted expression. Imprint establishment at hIC1 is impaired in the male germ line, which is associated with an abnormal composition of histone posttranslational modifications compared with mIC1. Overall, this study reveals evolutionarily divergent paternal imprinting at IC1 between mice and humans. The conserved maternal imprinting mechanism and function at IC1 demonstrates the possibility of modeling maternal transmission of hIC1 mutations associated with BWS in mice. In addition, we propose that further analyses in the paternal knock-in H19(+/hIC1) mice will elucidate the molecular mechanisms that may underlie SRS
Hutchins EK et al (2025), Supplementary Figures 1-6
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Supplemental Figure 1. Strain differences in retention of VitD metabolites after dietary depletion. (A) Percent retention of calcidiol and (B) calcitriol graphed by strain. Letters denote strains that differ by Tukey posthoc (a ≠ b = ab). Sample sizes: Total population (n=42) (HighC=24, LowC=18); n per strain (6); n per diet (21).
Supplemental Figure 2. Predictive relationship between canonical VitD activation genes and calcidiol and calcitriol under VDD conditions. (A) Association plots showing predictive relationship between Cyp2r1 or (B) Cyp27b1 transcript levels and serum calcidiol, calcitriol, or the activation ratio under VDD conditions only for all strains (left panel) or stratified by calcitriol phenotype (right panel). Activation ratio for one VDD-CC026 sample removed due to value >150x higher than other biological replicates. P-values, line of fit, and shaded 95% confidence interval are shown for each population tested. For significant associations (p<0.05), r2 indicates the amount of variance in metabolite concentrations explained by variance in Cyp2r1 or Cyp27b1. Sample sizes: Total population (n=41-42) (HighC=24, LowC=17-18); n per strain (5-6); n per diet (20-21).
Supplemental Figure 3. Genetic strain differences in Cyp27a1 expression. (A) Liver expression of Cyp24a1 (relative to Arpp0) graphed by strain or (B) calcitriol phenotype. P-values shown in (A) for strain (adjusted for diet), diet (adjusted for strain), and strain x diet effects. Sample sizes: Total population (n=42) (HighC=24, LowC=18); n per strain (6); n per diet (21).
Supplemental Figure 4. Genetic strain differences in Cubn expression. (A) Kidney expression of Cubn (relative to B2M) graphed by strain or (B) calcitriol phenotype. (C) Association plots showing relationship between Cubn and Lrp2 expression under VDS (left) or VDD (right) conditions. Measures for one VDD-CC006 outlier sample removed from (C) due to high variability from other biological replicates as depicted in (B). For (A), p-values shown for strain (adjusted for diet), diet (adjusted for strain), and strain x diet effects. For (C), p-values, line of fit, and shaded 95% confidence interval are shown for each population tested. For significant associations (p<0.05), r2 indicates the amount of variance explained. Sample sizes: Total population (n=41-42) (HighC=24, LowC=17-18); n per strain (5-6); n per diet (20-21).
Supplemental Figure 5. Western blots for renal VDR protein levels. Western blots showing bands specific for VDR and reference protein B-actin from mice under VDS (top panel) or VDD (bottom panel) conditions. Although blots were run simultaneously, sample order was reorganized for consistency throughout paper. Dashed vertical lines indicate where blot is discontinuous. Sample sizes: Total population (n=41) (HighC=24, LowC=18); n per strain (6); n per diet (21).
Supplemental Figure 6. Strain differences in serum calcium concentrations. Serum calcium concentrations graphed by strain (with samples from both diets combined since there was no diet effect). P-values shown for strain (adjusted for diet), diet (adjusted for strain), and strain x diet effects. Letters denote strains that differ by Tukey posthoc (a ≠ b = ab). Sample sizes: Total population (n=27) (HighC=15, LowC=12); n per strain (3-6); n per diet (13-14).
Hutchins EK et al (2025), Supplementary Table 1
No full textSupplementary Tables
Table 1. Descriptions of primers/oligos used for gene expression assays
Gestational diabetes mellitus placentas exhibit epimutations at placental development genes
No full textGestational diabetes mellitus (GDM) is a maternal metabolic disorder that perturbs placental development and increases the risk of offspring short- and long-term metabolic disorders. The mechanisms by which GDM impairs placental development remain poorly understood. Here, we defined the DNA methylome of GDM placentas and determined whether GDM perturbs methylation at genes important for placental development. We conducted an epigenome-wide association study of 42 placentas from pregnancies in the South African Soweto First 1000 days cohort (S1000). Using genome-wide bisulfite sequencing, we compared non-GDM placentas to GDM placentas with similar proportions from obese and non-obese mothers. Compared to non-GDM, GDM placentas exhibited a distinct methylation profile consisting of 12,210 differentially methylated CpGs (DMCs) that mapped to 3,875 genes. Epigenetically altered genes were enriched in Wnt and cadherin signalling pathways, both critical in placentation and embryogenesis. We also defined regional DNA methylation perturbation in GDM placentas at 11 placental development genes. These findings reveal extensive changes to the placental epigenome of GDM pregnancies and highlight perturbation enriched at important placental development genes. These molecular changes represent potential mechanisms for GDM-induced placental effects that may serve as candidate biomarkers for placental, maternal, and foetal health. Using a study design that used similar proportions of obese and non-obese mothers in our case and control pregnancies, we minimized the detection of changes due to obesity alone. Further work will be necessary to investigate the extent of the influence of obesity on these GDM-related placental epigenetic changes
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