Mediation Analysis Supports a Causal Relationship between Maternal Hyperglycemia and Placental DNA Methylation Variations at the Leptin Gene Locus and Cord Blood Leptin Levels
Abstract
:1. Introduction
2. Results
2.1. Sample Characteristics
2.2. Identification of Potential Functional LEP 5′-C-phosphate-G-3′ (CpG) Sites and Association with Childhood Anthropometric Profile
2.3. Investigation of Placental DNAm Levels as Mediating Factor between Maternal Glycemia and Adiposity Markers in Offspring
3. Methods
3.1. Participants’ Selection from Gen3G Cohort
3.2. Gen3G Follow-Up During Pregnancy
3.3. Sampling and Measures at Birth
3.4. Clinical Evaluation of Children
3.5. DNA Extraction and Sample Preparation
3.6. Measurements of DNA Methylation
3.7. Statistical Analyses
3.8. Mediation Analysis
4. Discussion
Strengths and Limitations
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
Abbreviations
BMI-z | Body mass index z-scores |
DNAm | DNA methylation |
CpG | 5′-C-phosphate-G-3′ |
DNMT | DNA methyltransferase |
DOHaD | Developmental origins of health and diseases |
GCT | Glucose challenge test |
GDM | Gestational diabetes mellitus |
Gen3G | Genetics of Glucose regulation in Gestation and Growth |
IQR | Interquartile range |
LEP | Leptin |
OGTT | Oral glucose tolerance test |
RCP | Regression on correlated probes |
RIN | RNA integrity number |
SS | Suprascapular |
ST | Skinfold Thickness |
TR | Tricipital |
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Participant Clinical Data n = 262 | Mean ± SD |
---|---|
Maternal characteristics | |
1st trimester of pregnancy | |
Age (years) | 28.6 ± 4.2 |
BMI (kg/m2) | 25.5 ± 5.5 |
Smoking during pregnancy | |
Yes | 7.4% |
No | 91.8% |
Glucose 1 h post 50 g GCT (mmol/l) | 5.0 ± 2.2 |
2nd trimester of pregnancy | |
Fasting Glucose (mmol/L) | 4.2 ± 0.3 |
Glucose 2 h post 75 g OGTT (mmol/L) | 5.9 ± 1.4 |
Child characteristics | |
At birth | |
Gestational age at birth (weeks) | 39.6 ± 1.0 |
Sex | |
Boys | 55% |
Girls | 45% |
Birthweight (kg) | 3.4 ± 0.4 |
Cord blood leptin levels (ng/mL) | 15,028 ± 12,791 |
At 3-years-old | |
Age (months) | 40.5 ± 3.0 |
Weight (kg) | 15.2 ± 1.7 |
BMI (kg/m2) | 16.1 ± 1.2 |
BMI z-scores | 0.5 ± 0.9 |
Sum of skinfolds thicknesses (mm)α | 17.5 ± 3.6 |
Ratio of skinfold thicknesses (SS:TR)α | 0.6 ± 0.1 |
CpG Sites from the MethylationEPIC BeadChip | Cord Blood Leptin Levels |
---|---|
cg15758240 | r = −0.289 p < 0.001 |
cg00011113 | r = −0.141 p = 0.03 |
cg05136031 | r = 0.221 p < 0.001 |
Mean for cg12782180, cg19594666, cg11045943, cg26814075, cg13381984 | r = −0.125 p = 0.05 |
cg00666422 | r = −0.129 p = 0.04 |
cg20564991 | r = −0.061 p = 0.34 |
cg23381058 | r = −0.198 p = 0.002 |
cg18603538 | r = 0.006 p = 0.93 |
cg04833007 | r = −0.045 p = 0.48 |
CpG Sites | BMI z-Scores | Sum of Skinfold Thickness | Ratio of Skinfold Thickness |
---|---|---|---|
cg15758240 | β = −0.391 p = 0.82 | β = 9.076 p = 0.21 | β = −0.581 p = 0.05 |
cg05136031 | β = −2.687 p = 0.05 | β = −1.441 p = 0.80 | β = −0.083 p = 0.72 |
cg23341058 | β = 0.019 p = 0.62 | β = 3.407 p = 0.35 | β = 0.015 p = 0.92 |
CpG Sites | Maternal Glucose 1 h Post 50 g GCT at 1st Trimester of Pregnancy | Maternal Fasting Glucose at 2nd Trimester of Pregnancy | Maternal Glucose 2 h Post 75 g OGTT at 2nd Trimester of Pregnancy |
---|---|---|---|
cg15758240 | β < 0.001 p = 0.74 | β = −0.013 p = 0.04 | β < 0.001 p = 0.21 |
cg05136031 | β < 0.001 p = 0.72 | Β < 0.001 p = 0.55 | Β < 0.001 p = 0.94 |
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Gagné-Ouellet, V.; Breton, E.; Thibeault, K.; Fortin, C.-A.; Cardenas, A.; Guérin, R.; Perron, P.; Hivert, M.-F.; Bouchard, L. Mediation Analysis Supports a Causal Relationship between Maternal Hyperglycemia and Placental DNA Methylation Variations at the Leptin Gene Locus and Cord Blood Leptin Levels. Int. J. Mol. Sci. 2020, 21, 329. https://doi.org/10.3390/ijms21010329
Gagné-Ouellet V, Breton E, Thibeault K, Fortin C-A, Cardenas A, Guérin R, Perron P, Hivert M-F, Bouchard L. Mediation Analysis Supports a Causal Relationship between Maternal Hyperglycemia and Placental DNA Methylation Variations at the Leptin Gene Locus and Cord Blood Leptin Levels. International Journal of Molecular Sciences. 2020; 21(1):329. https://doi.org/10.3390/ijms21010329
Chicago/Turabian StyleGagné-Ouellet, Valérie, Edith Breton, Kathrine Thibeault, Carol-Ann Fortin, Andres Cardenas, Renée Guérin, Patrice Perron, Marie-France Hivert, and Luigi Bouchard. 2020. "Mediation Analysis Supports a Causal Relationship between Maternal Hyperglycemia and Placental DNA Methylation Variations at the Leptin Gene Locus and Cord Blood Leptin Levels" International Journal of Molecular Sciences 21, no. 1: 329. https://doi.org/10.3390/ijms21010329