Quantification and Application of Potential Epigenetic Markers in Maternal Plasma of Pregnancies with Hypertensive Disorders
Abstract
:1. Introduction
2. Results and Discussion
2.1. Results
2.1.1. Clinical Characteristics
Characteristics | Controls (n = 188) | EO-PE (n = 16) | LO-PE (n = 47) | GH (n = 29) | P a | P b | P c | P d | P e | P f |
---|---|---|---|---|---|---|---|---|---|---|
Maternal age (years) | 33.0 (31.0–36.0) | 34.0 (31.6–36.0) | 34.1 (33.0–36.1) | 35.0 (33.0–37.0) | 0.204 | 0.061 | 0.059 | 0.383 | 0.393 | 0.809 |
BMI (kg/m2) | 20.2 (18.8–21.9) | 22.3 (22.1–29.4) | 22.6 (20.3–23.5) | 22.8 (21.2–30.5) | 0.000 | 0.000 | 0.000 | 0.073 | 0.948 | 0.044 |
Nulliparity | 76 (40.4) | 6 (37.5) | 24 (51.1) | 16 (55.2) | 0.819 | 0.187 | 0.135 | 0.397 | 0.353 | 0.815 |
Alcohol intake | 44 (23.4) | 6 (37.5) | 13 (27.7) | 5 (17.2) | 0.229 | 0.543 | 0.460 | 0.534 | 0.161 | 0.408 |
Tobacco use | 2 (1.1) | – | – | – | – | – | – | – | – | – |
Maximum SBP (mmHg) | 109.0 (101.0–118.0) | 162.0 (160.0–173.0) | 147.0 (140.5–150.5) | 157.0 (150.5–167.0) | 0.000 | 0.000 | 0.000 | 0.000 | 0.387 | 0.000 |
Maximum DBP (mmHg) | 63.0 (57.0–69.0) | 99.0 (93.0–100.0) | 90.0 (86.0–94.0) | 96.0 (89.0–98.5) | 0.000 | 0.000 | 0.000 | 0.035 | 0.355 | 0.077 |
Birthweight (g) | 3290.0 (3040.0–3495.0) | 2265.0 (1878.8–2362.5) | 2920.0 (2572.5–3297.5) | 3012.5 (2800.0–3290.0) | 0.000 | 0.000 | 0.001 | 0.002 | 0.000 | 0.384 |
Sex ratio of fetus (male:female) | 92:96 | 10:6 | 16:31 | 11:18 | 0.298 | 0.067 | 0.269 | 0.076 | 0.133 | 0.807 |
GA at delivery (weeks) | 39.5 (39.1–40.3) | 35.8 (34.4–36.4) | 38.3 (36.8–39.3) | 38.6 (38.2–39.5) | 0.000 | 0.000 | 0.006 | 0.003 | 0.000 | 0.032 |
GA at sampling (weeks) | – | – | – | – | – | – | – | – | – | – |
First trimester | 12.1 (8.4–12.4) | 12.0 (11.1–12.3) | 12.3 (11.7–12.5) | 12.3 (12.2–12.3) | 0.850 | 0.382 | 0.205 | 0.571 | 0.567 | 0.970 |
Sceond trimester | 21.2 (16.4–25.0) | 20.9 (16.6–25.2) | 18.0 (16.6–24.0) | 17.9 (16.3–24.4) | 0.722 | 0.060 | 0.090 | 0.451 | 0.189 | 0.903 |
Third trimester | 36.5 (36.1–37.0) | 36.3 (35.3–36.6) | 36.9 (36.2–38.2) | 36.6 (36.6–37.7) | 0.063 | 0.329 | 0.100 | 0.187 | 0.073 | 0.967 |
2.1.2. Selection of Fetal-Specific Hypermethylated CpG loci by High-resolution Tiling Array Analysis
Target Regions | Chr. | Description | Probe Position | Placenta (Log2) | Normal Blood (Log2) | ||
---|---|---|---|---|---|---|---|
Start | Stop | Pregnant | Non-Pregnant | ||||
SOD1 | 21 | Inside | 33037889 | 33037933 | 2.62 ± 0.53 | 0.00 ± 0.00 | 0.00 ± 0.00 |
DSCR3 | 21 | Inside | 38629500 | 38629559 | 2.77 ± 0.43 | 0.00 ± 0.00 | 0.00 ± 0.00 |
C2CD2 | 21 | Inside | 43316160 | 43316204 | 3.45 ± 1.02 | 0.00 ± 0.00 | 0.00 ± 0.00 |
UMODL1 | 21 | Inside | 43483898 | 43483942 | 3.14 ± 0.84 | 0.00 ± 0.00 | 0.00 ± 0.00 |
ENST00000433952: -7924 | 21 | Promoter | 40357602 | 40357646 | 3.39 ± 0.77 | 0.00 ± 0.00 | 0.00 ± 0.00 |
ENST00000433952: -7988 | 21 | Promoter | 40357666 | 40357710 | 3.52 ± 1.12 | 0.00 ± 0.00 | 0.00 ± 0.00 |
ENST00000450830: -4439 | 21 | Promoter | 34524477 | 34524521 | 2.85 ± 0.82 | 0.00 ± 0.00 | 0.00 ± 0.00 |
2.1.3. Confirmation of High-Resolution Tiling Array Data Using Bisulfite Direct Sequencing
2.1.4. Quantification of DSCR3, RASSF1A, SRY and GADH in Maternal Plasma of Normal Pregnancies
2.1.5. DSCR3 and RASSF1A Concentrations in Maternal Plasma between Normal Pregnancies and Pregnancies with Hypertensive Disorders
2.2. Discussion
Marker | Controls | EO-PE | LO-PE | GH | P a | P b | P c | P d | P e | P f |
---|---|---|---|---|---|---|---|---|---|---|
6–14 weeks | (n = 52) | (n = 4) | (n = 10) | (n = 10) | – | – | – | – | – | – |
DSCR3 | 890.9 (538.9–1530.0) | 1007.7 (721.5–1454.3) | 939.1 (699.6–1718.2) | 1098.3 (561.7–2073.6) | 0.818 | 0.467 | 0.566 | 0.945 | 0.733 | 0.739 |
RASSF1A | 445.6 (262.9–936.4) | 858.0 (652.6–1215.2) | 595.2 (382.1–807.2) | 450.1 (385.1–719.7) | 0.118 | 0.503 | 0.444 | 0.374 | 0.188 | 0.853 |
15–23 weeks | (n = 31) | (n = 3) | (n = 16) | (n = 7) | – | – | – | – | – | – |
DSCR3 | 1779.7 (1443.8–2590.6) | 2594.0 (2171.5–3075.6) | 2573.7 (2159.9–2936.4) | 1692.8 (1220.7–2009.1) | 0.289 | 0.121 | 0.580 | 0.958 | 0.183 | 0.076 |
RASSF1A | 1333.8 (727.9–2403.3) | 2305.5 (1571.0–2842.2) | 1601.4 (1154.4–2036.8) | 1013.3 (716.5–1375.0) | 0.449 | 0.946 | 0.282 | 0.421 | 0.183 | 0.103 |
24–32 weeks | (n = 47) | (n = 5) | (n = 7) | (n = 5) | – | – | – | – | – | – |
DSCR3 | 2521.3 (1824.9–4438.2) | 6772.5 (5251.1–7876.2) | 3295.0 (1118.3–6765.2) | 2725.5 (1372.9–6415.1) | 0.018 | 0.920 | 0.857 | 0.268 | 0.222 | 1.000 |
RASSF1A | 1228.3 (827.6–2357.1) | 3100.8 (2359.7–4600.0) | 819.1 (610.1–1177.3) | 1765.5 (406.2–2965.0) | 0.032 | 0.212 | 1.000 | 0.149 | 0.222 | 0.639 |
33–41 weeks | (n = 58) | (n = 4) | (n = 14) | (n = 7) | – | – | – | – | – | – |
DSCR3 | 4212.9 (2841.1–6162.5) | 10,632.4 (8426.2–16,027.7) | 11,164.3 (7733.5–13,909.9) | 6985.4 (5070.0–8881.8) | 0.001 | 0.000 | 0.079 | 0.721 | 0.109 | 0.056 |
RASSF1A | 1777.2 (930.2–2674.1) | 4002.8 (3449.6–5142.4) | 5051.3 (3441.6–6787.2) | 2979.7 (1915.7–3763.8) | 0.008 | 0.000 | 0.173 | 0.721 | 0.109 | 0.016 |
3. Materials and Methods
3.1. Study Participants and Samples
3.2. DNA Preparation
3.3. Data Analysis of High-Resolution Tiling Arrays
3.4. Bisulfite Direct Sequencing
3.5. Methylation Quantification of Bisulfite Direct Sequencing Data
3.6. Enrichment of Methylated DNA
3.7. Quantitative Real-Time PCR
3.8. Statistical Analysis
4. Conclusions
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Kim, H.J.; Kim, S.Y.; Lim, J.H.; Kwak, D.W.; Park, S.Y.; Ryu, H.M. Quantification and Application of Potential Epigenetic Markers in Maternal Plasma of Pregnancies with Hypertensive Disorders. Int. J. Mol. Sci. 2015, 16, 29875-29888. https://doi.org/10.3390/ijms161226201
Kim HJ, Kim SY, Lim JH, Kwak DW, Park SY, Ryu HM. Quantification and Application of Potential Epigenetic Markers in Maternal Plasma of Pregnancies with Hypertensive Disorders. International Journal of Molecular Sciences. 2015; 16(12):29875-29888. https://doi.org/10.3390/ijms161226201
Chicago/Turabian StyleKim, Hyun Jin, Shin Young Kim, Ji Hyae Lim, Dong Wook Kwak, So Yeon Park, and Hyun Mee Ryu. 2015. "Quantification and Application of Potential Epigenetic Markers in Maternal Plasma of Pregnancies with Hypertensive Disorders" International Journal of Molecular Sciences 16, no. 12: 29875-29888. https://doi.org/10.3390/ijms161226201