Prenatal Synthetic Glucocorticoid Treatment Changes DNA Methylation States in Male Organ Systems: Multigenerational Effects

Prenatal synthetic glucocorticoids (sGC) are administered to pregnant women at risk of delivering preterm, approximately 10% of all pregnancies. Animal studies have demonstrated that offspring exposed to elevated glucocorticoids, either by administration of sGC or as a result of maternal stress, are at increased risk of developing behavioral, endocrine, and metabolic abnormalities. DNA methylation is a covalent modification of DNA that plays a critical role in long-lasting programming of gene expression. Here we tested the hypothesis that prenatal sGC treatment has both acute and long-term effects on DNA methylation states in the fetus and offspring and that these effects extend into a subsequent generation. Pregnant guinea pigs were treated with sGC in late gestation, and methylation analysis by luminometric methylation assay was undertaken in organs from fetuses and offspring across two generations. Expression of genes that modify the epigenetic state were measured by quantitative real-time PCR. Results indicate that there are organ-specific developmental trajectories of methylation in the fetus and newborn. Furthermore, these trajectories are substantially modified by intrauterine exposure to sGC. These sGC-induced changes in DNA methylation remain into adulthood and are evident in the next generation. Furthermore, prenatal sGC exposure alters the expression of several genes encoding proteins that modulate the epigenetic state. Several of these changes are long lasting and are also present in the next generation. These data support the hypothesis that prenatal sGC exposure leads to broad changes in critical components of the epigenetic machinery and that these effects can pass to the next generation.

Download Full-text

Glucocorticoid Programming of the Fetal Male Hippocampal Epigenome

Endocrinology ◽

10.1210/en.2012-1980 ◽

2013 ◽

Vol 154 (3) ◽

pp. 1168-1180 ◽

Cited By ~ 62

Author(s):

Ariann Crudo ◽

Matthew Suderman ◽

Vasilis G. Moisiadis ◽

Sophie Petropoulos ◽

Alisa Kostaki ◽

...

Keyword(s):

Dna Methylation ◽

Promoter Methylation ◽

Plasma Cortisol ◽

Late Gestation ◽

Gene Promoters ◽

Fetal Plasma ◽

Genome Wide ◽

Organ Systems ◽

Increased Risk ◽

H3k9 Acetylation

Abstract The late-gestation surge in fetal plasma cortisol is critical for maturation of fetal organ systems. As a result, synthetic glucocorticoids (sGCs) are administered to pregnant women at risk of delivering preterm. However, animal studies have shown that fetal exposure to sGC results in increased risk of behavioral, endocrine, and metabolic abnormalities in offspring. Here, we test the hypothesis that prenatal GC exposure resulting from the fetal cortisol surge or after sGC exposure results in promoter-specific epigenetic changes in the hippocampus. Fetal guinea pig hippocampi were collected before (gestational day [GD52]) and after (GD65) the fetal plasma cortisol surge (Term∼GD67) and 24 hours after (GD52) and 14 days after (GD65) two repeat courses of maternal sGC (betamethasone) treatment (n = 3–4/gp). We identified extensive genome-wide alterations in promoter methylation in late fetal development (coincident with the fetal cortisol surge), whereby the majority of the affected promoters exhibited hypomethylation. Fetuses exposed to sGC in late gestation exhibited substantial differences in DNA methylation and histone h3 lysine 9 (H3K9) acetylation in specific gene promoters; 24 hours after the sGC treatment, the majority of genes affected were hypomethylated or hyperacetylated. However, 14 days after sGC exposure these differences did not persist, whereas other promoters became hypermethylated or hyperacetylated. These data support the hypothesis that the fetal GC surge is responsible, in part, for significant variations in genome-wide promoter methylation and that prenatal sGC treatment profoundly changes the epigenetic landscape, affecting both DNA methylation and H3K9 acetylation. This is important given the widespread use of sGC in the management of women in preterm labor.

Download Full-text

DNA methylome signatures of prenatal exposure to synthetic glucocorticoids in hippocampus and peripheral whole blood of female guinea pigs in early life

Translational Psychiatry ◽

10.1038/s41398-020-01186-6 ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Aya Sasaki ◽

Margaret E. Eng ◽

Abigail H. Lee ◽

Alisa Kostaki ◽

Stephen G. Matthews

Keyword(s):

Dna Methylation ◽

Whole Blood ◽

Guinea Pigs ◽

Critical Role ◽

Methylation Status ◽

Peripheral Tissues ◽

Epigenetic Biomarkers ◽

Differentially Methylated Genes ◽

Increased Risk ◽

Synthetic Glucocorticoids

AbstractSynthetic glucocorticoids (sGC) are administered to women at risk of preterm delivery, approximately 10% of all pregnancies. In animal models, offspring exposed to elevated glucocorticoids, either by administration of sGC or endogenous glucocorticoids as a result of maternal stress, show increased risk of developing behavioral, endocrine, and metabolic dysregulation. DNA methylation may play a critical role in long-lasting programming of gene regulation underlying these phenotypes. However, peripheral tissues such as blood are often the only accessible source of DNA for epigenetic analyses in humans. Here, we examined the hypothesis that prenatal sGC administration alters DNA methylation signatures in guinea pig offspring hippocampus and whole blood. We compared these signatures across the two tissue types to assess epigenetic biomarkers of common molecular pathways affected by sGC exposure. Guinea pigs were treated with sGC or saline in late gestation. Genome-wide modifications of DNA methylation were analyzed at single nucleotide resolution using reduced representation bisulfite sequencing in juvenile female offspring. Results indicate that there are tissue-specific as well as common methylation signatures of prenatal sGC exposure. Over 90% of the common methylation signatures associated with sGC exposure showed the same directionality of change in methylation. Among differentially methylated genes, 134 were modified in both hippocampus and blood, of which 61 showed methylation changes at identical CpG sites. Gene pathway analyses indicated that prenatal sGC exposure alters the methylation status of gene clusters involved in brain development. These data indicate concordance across tissues of epigenetic programming in response to alterations in glucocorticoid signaling.

Download Full-text

Effects of Antenatal Synthetic Glucocorticoid on Glucocorticoid Receptor Binding, DNA Methylation, and Genome-Wide mRNA Levels in the Fetal Male Hippocampus

Endocrinology ◽

10.1210/en.2013-1484 ◽

2013 ◽

Vol 154 (11) ◽

pp. 4170-4181 ◽

Cited By ~ 46

Author(s):

Ariann Crudo ◽

Sophie Petropoulos ◽

Matthew Suderman ◽

Vasilis G. Moisiadis ◽

Alisa Kostaki ◽

...

Keyword(s):

Dna Methylation ◽

Dna Binding ◽

Glucocorticoid Receptor ◽

Fetal Lung ◽

Late Gestation ◽

Mrna Levels ◽

Substantial Impact ◽

Genome Wide ◽

Organ Systems ◽

Endogenous Cortisol

The endogenous glucocorticoid (GC) surge in late gestation plays a vital role in maturation of several organ systems. For this reason, pregnant women at risk of preterm labor are administered synthetic glucocorticoids (sGCs) to promote fetal lung development. Animal studies have shown that fetal sGC exposure can cause life-long changes in endocrine and metabolic function. We have previously shown that antenatal sGC treatment is associated with alterations in global DNA methylation and modifications to the hippocampal methylome and acetylome. In this study, we hypothesized that: 1) there are changes in the transcriptional landscape of the fetal hippocampus in late gestation, associated with the endogenous cortisol surge; 2) fetal sGC exposure alters genome-wide transcription in the hippocampus; and 3) these changes in transcription are associated with modified glucocorticoid receptor (GR) DNA binding and DNA methylation. sGC was administered as 2 courses on gestational days (GD) 40, 41, 50, and 51, and the hippocampi of fetal guinea pigs were examined before (GD52) and after (GD65) the endogenous cortisol surge (Term ∼GD67). We also analyzed fetal hippocampi 24 hours and 14 days following maternal sGC injections (n = 3–4/group). Genome-wide modification of transcription and GR DNA binding occurred in late gestation, in parallel with the normal GC surge. Further, sGC exposure had a substantial impact on the hippocampal transcriptome, GR-DNA binding, and DNA methylation at 24 hours and 14 days following the final sGC treatment. These data support the hypothesis that GC exposure in late gestation plays a significant role in modifying the transcriptional and epigenetic landscape of the developing fetal hippocampus and that substantial effects are evident for at least 2 weeks after sGC exposure.

Download Full-text

Understanding the effects of maternal undernutrition in early gestation on placental development in late gestation

10.32469/10355/67713 ◽

2018 ◽

Author(s):

◽

Gerialisa Allison Marcella Van Gronigen Case

Keyword(s):

Cardiovascular Disease ◽

Dna Methylation ◽

Maternal Nutrition ◽

Financial Burden ◽

Late Gestation ◽

Placental Development ◽

Early Gestation ◽

Maternal Undernutrition ◽

Increased Risk ◽

The U.S

Chronic diseases such as cardiovascular disease and diabetes are on the rise among the U.S. population. Heart Disease is the leading cause of death in the US and is responsible 610,000 deaths per year. A suite of risk factors for cardiovascular disease and diabetes â€" obesity, elevated blood sugar,hypertension, excess cholesterol, and elevated triglycerides, referred to asMetabolic Syndrome, contributes to an increased risk of mortality. These too areincreasingly prevalent. For example, according to the CDC, in 2011 more than 35% of the U.S. population was obese, with a financial burden of US $147 billion annually. Though improper diet, lack of exercise and socioeconomic status areassociated with these adult diseases, inadequate maternal nutrition just prior toor during pregnancy is also a risk factor. The goal of my research is to identifythe critical time point by which maternal under-nutrition (MUN) during earlygestation permanently alters placental development, the mechanism by whichthis occurs, and whether leptin acts as a mediator.The relationship between the maternal environment and long-term healthof offspring is referred to as the developmental origins of health and disease (DOHaD), or sometimes fetal programming. MUN during pregnancy may affectdeveloping organs to alter the phenotypic outcome of the offspring to promoteadult disease. The timing of the insult is also critical. During the periconceptionalperiod and early gestation, there are major developmental processes such asembryonic organogenesis, and placenta formation. There are also two rounds of DNA methylation reprogramming that occur. I investigated the effects of maternalunder-nutrition from three weeks prior to gestation to d 11.5 (mid gestation inmice). I found that maternal periconceptional food restriction until mid- gestationaffected formation of blood spaces within the labyrinthine placenta near term, ascompared to those from control-fed dams. There was also a three-fold decreasein mRNA encoding SNAT4, an amino acid transporter, in restricted placentae.These changes in placentae from dams fed a nutrient restricted diet suggestsome compensation to ensure adequate nutrients are available to the fetus forproper development.I also examined the effects MUN undernutrition during the first half ofgestation had on placental DNA methylation at late gestation and whether leptinwould act as a mediator of these effects. It is not entirely understood howinformation about maternal nutrition is sensed by the placenta. Leptin, is ahormone that is secreted by fat tissue and plays a role in energy homeostasis,metabolism and reproduction. Therefore, it could be one potential mechanism bywhich maternal nutrition can influence placental function and in turn, fetaldevelopment. I found placentae from dams who experienced MUN and leptinsupplementation had a greater number of differentially methylated regionscompared to placentae from dams fed an adequate fed diet than did placentae ofdams fed MUN only. Regions that were differentially methylated by maternal dietand/or leptin supplementation were located near genes in pathways involved inmolecular transport and carbohydrate metabolism as well as postnatal growth,weight and body size, formation of new blood vessels during embryogenesis. These pathways suggest a link between maternal food deprivation andoffspring's' growth, metabolism and formation of the cardiovascular system.

Download Full-text

ANTENATAL GLUCOCORTICOIDS: IS THERE CAUSE FOR CONCERN?

Fetal and Maternal Medicine Review ◽

10.1017/s0965539503001141 ◽

2003 ◽

Vol 14 (4) ◽

pp. 329-354 ◽

Cited By ~ 12

Author(s):

MARCUS H ANDREWS ◽

STEPHEN G MATTHEWS

Keyword(s):

Controlled Trial ◽

Intraventricular Haemorrhage ◽

Glucocorticoid Therapy ◽

Long Term Effects ◽

Glucocorticoid Treatment ◽

Organ Systems ◽

Term Labour ◽

The Brain ◽

Antenatal Treatment

Antenatal glucocorticoid therapy was pioneered by Liggins & Howie over three decades ago. In a controlled trial of babies delivering before 32 weeks gestation, it was shown that antenatal glucocorticoid treatment reduced the incidence of respiratory distress syndrome (RDS) and significantly lowered neonatal mortality. Synthetic glucocorticoid (sGC) therapy is also associated with reduced risk of neonatal intraventricular haemorrhage, necrotizing entercolitis and hyperbilirubinaemia. The benefit of antenatal glucocorticoid therapy to lung function in the newborn infant has led to routine use in situations of threatened pre-term labour. In 1994, a National Institute of Health Consensus Development Conference recommended antenatal treatment of all women at risk of preterm delivery, between 24 and 34 weeks of gestation, with sGC. Similar statements were issued in Europe, Canada and Australia. However, the NIH Consensus Report highlighted the requirement for further research into the long-term effects of antenatal sGC treatment on development of the brain and other organ systems, and their function after birth.

Download Full-text

Opposing Epigenetic Signatures in Human Sperm by Intake of Fast Food Versus Healthy Food

Frontiers in Endocrinology ◽

10.3389/fendo.2021.625204 ◽

2021 ◽

Vol 12 ◽

Author(s):

Adelheid Soubry ◽

Susan K. Murphy ◽

Greet Vansant ◽

Yang He ◽

Thomas M. Price ◽

...

Keyword(s):

Dna Methylation ◽

Chronic Diseases ◽

Fast Food ◽

Dietary Habits ◽

Imprinted Genes ◽

Sperm Cells ◽

Next Generation ◽

Eating Pattern ◽

Food Items ◽

Increased Risk

Animal experiments have demonstrated that diets high in fats create a harmful environment for developing sperm cells, contributing to impaired reproductive health and induced risk for chronic diseases in the next generation. Changes at the level of the epigenome have been suggested to underlie these observations. Human data are limited to verify this hypothesis. While we earlier demonstrated a link between male obesity and DNA methylation changes at imprinted genes in mature sperm cells and newborns, it is currently unknown if -or how- a paternal eating pattern (related to obesity) is related to indices for epigenetic inheritance. We here aim to examine a yet unexplored link between consumption of healthy (rich in vitamins and fibers) or unhealthy (“fast”) foods and methylation at imprint regulatory regions in DNA of sperm. We obtained semen and data from 67 men, as part of a North Carolina-based study: The Influence of the Environment on Gametic Epigenetic Reprogramming (TIEGER) study. Dietary data included intake of fruits/nuts, vegetables/soups, whole grain bread, meat, seafood/fish, and fatty or processed food items. Multiple regression models were used to explore the association between dietary habits and clinical sperm parameters as well as DNA methylation levels, quantified using bisulfite pyrosequencing at 12 differentially methylated regions (DMRs) of the following imprinted genes: GRB10, IGF2, H19, MEG3, NDN, NNAT, PEG1/MEST, PEG3, PLAGL1, SNRPN, and SGCE/PEG10. After adjusting for age, obesity status and recruitment method, we found that Total Motile Count (TMC) was significantly higher if men consumed fruits/nuts (β=+6.9, SE=1.9, p=0.0005) and vegetables (β=+5.4, SE=1.9, p=0.006), whereas consumption of fries was associated with lower TMC (β=-20.2, SE=8.7, p=0.024). Semen volume was also higher if vegetables or fruits/nuts were frequently consumed (β=+0.06, SE=0.03, p=0.03). Similarly, our sperm epigenetic analyses showed opposing associations for healthy versus fast food items. Frequent consumption of fries was related to a higher chance of sperm being methylated at the MEG3-IG CpG4 site (OR=1.073, 95%CI: 1.035-1.112), and high consumption of vegetables was associated with a lower risk of DNA methylation at the NNAT CpG3 site (OR=0.941, 95%CI: 0.914-0.968). These results remained significant after adjusting for multiple testing. We conclude that dietary habits are linked to sperm epigenetic outcomes. If carried into the next generation paternal unhealthy dietary patterns may result in adverse metabolic conditions and increased risk for chronic diseases in offspring.

Download Full-text

Profile of copper-associated DNA methylation and its association with incident acute coronary syndrome

Clinical Epigenetics ◽

10.1186/s13148-021-01004-w ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Pinpin Long ◽

Qiuhong Wang ◽

Yizhi Zhang ◽

Xiaoyan Zhu ◽

Kuai Yu ◽

...

Keyword(s):

Dna Methylation ◽

Acute Coronary Syndrome ◽

Methylation Level ◽

Meta Analysis ◽

Regulation Of Gene Expression ◽

Density Lipoprotein ◽

Blood Leukocytes ◽

Coronary Syndrome ◽

A Genome ◽

Increased Risk

Abstract Background Acute coronary syndrome (ACS) is a cardiac emergency with high mortality. Exposure to high copper (Cu) concentration has been linked to ACS. However, whether DNA methylation contributes to the association between Cu and ACS is unclear. Methods We measured methylation level at > 485,000 cytosine-phosphoguanine sites (CpGs) of blood leukocytes using Human Methylation 450 Bead Chip and conducted a genome-wide meta-analysis of plasma Cu in a total of 1243 Chinese individuals. For plasma Cu-related CpGs, we evaluated their associations with the expression of nearby genes as well as major cardiovascular risk factors. Furthermore, we examined their longitudinal associations with incident ACS in the nested case-control study. Results We identified four novel Cu-associated CpGs (cg20995564, cg18608055, cg26470501 and cg05825244) within a 5% false discovery rate (FDR). DNA methylation level of cg18608055, cg26470501, and cg05825244 also showed significant correlations with expressions of SBNO2, BCL3, and EBF4 gene, respectively. Higher DNA methylation level at cg05825244 locus was associated with lower high-density lipoprotein cholesterol level and higher C-reactive protein level. Furthermore, we demonstrated that higher cg05825244 methylation level was associated with increased risk of ACS (odds ratio [OR], 1.23; 95% CI 1.02–1.48; P = 0.03). Conclusions We identified novel DNA methylation alterations associated with plasma Cu in Chinese populations and linked these loci to risk of ACS, providing new insights into the regulation of gene expression by Cu-related DNA methylation and suggesting a role for DNA methylation in the association between copper and ACS.

Download Full-text

Methamphetamine use and psychotic symptoms: findings from a New Zealand longitudinal birth cohort

Psychological Medicine ◽

10.1017/s0033291721002415 ◽

2021 ◽

pp. 1-8

Author(s):

Joseph M. Boden ◽

James A. Foulds ◽

Giles Newton-Howes ◽

Rebecca McKetin

Keyword(s):

New Zealand ◽

General Population ◽

Birth Cohort ◽

Individual Characteristics ◽

Psychotic Symptoms ◽

Confounding Factors ◽

Long Term Effects ◽

Methamphetamine Use ◽

Increased Risk ◽

Psychosis Risk

Abstract Background This study examined the association between methamphetamine use and psychotic symptoms in a New Zealand general population birth cohort (n = 1265 at birth). Methods At age 18, 21, 25, 30, and 35, participants reported on their methamphetamine use and psychotic symptoms in the period since the previous interview. Generalized estimating equations modelled the association between methamphetamine use and psychotic symptoms (percentage reporting any symptom, and number of symptoms per participant). Confounding factors included childhood individual characteristics, family socioeconomic circumstances and family functioning. Long term effects of methamphetamine use on psychotic symptoms were assessed by comparing the incidence of psychotic symptoms at age 30–35 for those with and without a history of methamphetamine use prior to age 30. Results After adjusting for confounding factors and time-varying covariate factors including concurrent cannabis use, methamphetamine use was associated with a modest increase in psychosis risk over five waves of data (adjusted odds ratio (OR) 1.33, 95% confidence interval (CI) 1.03–1.72 for the percentage measure; and IRR 1.24, 95% CI 1.02–1.50 for the symptom count measure). The increased risk of psychotic symptoms was concentrated among participants who had used at least weekly at any point (adjusted OR 2.85, 95% CI 1.21–6.69). Use of methamphetamine less than weekly was not associated with increased psychosis risk. We found no evidence for a persistent vulnerability to psychosis in the absence of continuing methamphetamine use. Conclusion Methamphetamine use is associated with increased risk of psychotic symptoms in the general population. Increased risk is chiefly confined to people who ever used regularly (at least weekly), and recently.

Download Full-text

Epigenetic Contribution and Genomic Imprinting Dlk1-Dio3 miRNAs in Systemic Lupus Erythematosus

Genes ◽

10.3390/genes12050680 ◽

2021 ◽

Vol 12 (5) ◽

pp. 680

Author(s):

Rujuan Dai ◽

Zhuang Wang ◽

S. Ansar Ahmed

Keyword(s):

Systemic Lupus Erythematosus ◽

Dna Methylation ◽

Lupus Erythematosus ◽

Critical Role ◽

Methylation Status ◽

Transcriptional Level ◽

Dna Hypomethylation ◽

Systemic Lupus ◽

Multiple Organs ◽

Genetic Imprinting

Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease that afflicts multiple organs, especially kidneys and joints. In addition to genetic predisposition, it is now evident that DNA methylation and microRNAs (miRNAs), the two major epigenetic modifications, are critically involved in the pathogenesis of SLE. DNA methylation regulates promoter accessibility and gene expression at the transcriptional level by adding a methyl group to 5′ cytosine within a CpG dinucleotide. Extensive evidence now supports the importance of DNA hypomethylation in SLE etiology. miRNAs are small, non-protein coding RNAs that play a critical role in the regulation of genome expression. Various studies have identified the signature lupus-related miRNAs and their functional contribution to lupus incidence and progression. In this review, the mutual interaction between DNA methylation and miRNAs regulation in SLE is discussed. Some lupus-associated miRNAs regulate DNA methylation status by targeting the DNA methylation enzymes or methylation pathway-related proteins. On the other hand, DNA hyper- and hypo-methylation are linked with dysregulated miRNAs expression in lupus. Further, we specifically discuss the genetic imprinting Dlk1-Dio3 miRNAs that are subjected to DNA methylation regulation and are dysregulated in several autoimmune diseases, including SLE.

Download Full-text

Epigenome-wide association study of whole blood gene expression in Framingham Heart Study participants provides molecular insight into the potential role of CHRNA5 in cigarette smoking-related lung diseases

Clinical Epigenetics ◽

10.1186/s13148-021-01041-5 ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Chen Yao ◽

Roby Joehanes ◽

Rory Wilson ◽

Toshiko Tanaka ◽

Luigi Ferrucci ◽

...

Keyword(s):

Gene Expression ◽

Lung Cancer ◽

Dna Methylation ◽

Cigarette Smoking ◽

Association Study ◽

Whole Blood ◽

Lung Diseases ◽

Epigenetic Modification ◽

Blood Gene Expression ◽

Increased Risk

Abstract Background DNA methylation is a key epigenetic modification that can directly affect gene regulation. DNA methylation is highly influenced by environmental factors such as cigarette smoking, which is causally related to chronic obstructive pulmonary disease (COPD) and lung cancer. To date, there have been few large-scale, combined analyses of DNA methylation and gene expression and their interrelations with lung diseases. Results We performed an epigenome-wide association study of whole blood gene expression in ~ 6000 individuals from four cohorts. We discovered and replicated numerous CpGs associated with the expression of cis genes within 500 kb of each CpG, with 148 to 1,741 cis CpG-transcript pairs identified across cohorts. We found that the closer a CpG resided to a transcription start site, the larger its effect size, and that 36% of cis CpG-transcript pairs share the same causal genetic variant. Mendelian randomization analyses revealed that hypomethylation and lower expression of CHRNA5, which encodes a smoking-related nicotinic receptor, are causally linked to increased risk of COPD and lung cancer. This putatively causal relationship was further validated in lung tissue data. Conclusions Our results provide a large and comprehensive association study of whole blood DNA methylation with gene expression. Expression platform differences rather than population differences are critical to the replication of cis CpG-transcript pairs. The low reproducibility of trans CpG-transcript pairs suggests that DNA methylation regulates nearby rather than remote gene expression. The putatively causal roles of methylation and expression of CHRNA5 in relation to COPD and lung cancer provide evidence for a mechanistic link between patterns of smoking-related epigenetic variation and lung diseases, and highlight potential therapeutic targets for lung diseases and smoking cessation.

Download Full-text