Visceral Adipose Tissue Inflammatory Factors (TNF-Alpha, SOCS3) in Gestational Diabetes (GDM): Epigenetics as a Clue in GDM Pathophysiology

Rebecca C Rancourt, Raffael Ott, Thomas Ziska, Karen Schellong, Kerstin Melchior, Wolfgang Henrich, Andreas Plagemann, Rebecca C Rancourt, Raffael Ott, Thomas Ziska, Karen Schellong, Kerstin Melchior, Wolfgang Henrich, Andreas Plagemann

Abstract

Gestational diabetes (GDM) is among the most challenging diseases in westernized countries, affecting mother and child, immediately and in later life. Obesity is a major risk factor for GDM. However, the impact visceral obesity and related epigenetics play for GDM etiopathogenesis have hardly been considered so far. Our recent findings within the prospective 'EaCH' cohort study of women with GDM or normal glucose tolerance (NGT), showed the role, critical factors of insulin resistance (i.e., adiponectin, insulin receptor) may have for GDM pathophysiology with epigenetically modified expression in subcutaneous (SAT) and visceral (VAT) adipose tissues. Here we investigated the expression and promoter methylation of key inflammatory candidates, tumor necrosis factor-alpha (TNF-α) and suppressor of cytokine signaling 3 (SOCS3) in maternal adipose tissues collected during caesarian section (GDM, n = 19; NGT, n = 22). The mRNA expression of TNF-α and SOCS3 was significantly increased in VAT, but not in SAT, of GDM patients vs. NGT, accompanied by specific alterations of respective promoter methylation patterns. In conclusion, we propose a critical role of VAT and visceral obesity for the pathogenesis of GDM, with epigenetic alterations of the expression of inflammatory factors as a potential factor.

Keywords: DNA methylation; SOCS3; TNF-α; adipose tissue; epigenetics; gestational diabetes mellitus; mRNA expression.

Conflict of interest statement

The authors declare that they have no competing interests.

Figures

Figure 1
Figure 1
Relative mRNA levels of TNF-α and suppressor of cytokine signaling 3 (SOCS3) in subcutaneous adipose tissue (SAT) and visceral adipose tissue (VAT) of women with GDM vs. NGT. Relative gene expression of TNF-α (A) and SOCS3 (B) was normalized to peptidylprolyl isomerase A (PPIA) in abdominal SAT and omental VAT, of women with GDM (n = 19, black) vs. NGT women (n = 22, white). Data are means ± SEM, shown as percentage to NGT levels. A.U., arbitrary units. TNF-α VAT ** P < 0.0001, SOCS3-VAT * P = 0.01. (C–D) Pearson’s correlation coefficients (R) calculating the relationship between maternal blood TNF-α levels and VAT gene expression data. NGT: open circles, GDM: red circles. Statistical significance between groups (A,B) and for correlations (C,D) remained after adjustment for prepregnancy body-mass-index (BMI) and BMI at delivery.
Figure 2
Figure 2
DNA methylation analysis in the TNF-α promoter region. CpG site-specific DNA methylation analyses at the TNF-α promoter region in visceral adipose tissue from mothers with NGT vs. GDM. (A) Schematic illustration of the DNA methylation assays (R1, R2, and R3) for the TNF-α promoter region, including transcription factor binding sites (TFBS) Sp1, EGR1, AP2 (diamonds). (B) Percent DNA methylation at each individual CpG site investigated (10 CpG sites) in VAT of normal glucose tolerant (NGT; white; n = 22) vs. GDM group (GDM; black; n = 19). Overall mean across CpG sites is also included. Data are means ± S.E.M. * P < 0.05. TSS: Transcriptional start site, bp: basepairs.
Figure 3
Figure 3
DNA methylation analysis in the SOCS3 promoter region. CpG site-specific DNA methylation analyses at SOCS3 within the promoter region in visceral adipose tissue from mothers with NGT vs. GDM. (A) Schematic illustration of the DNA methylation assays (R1, R2, and R3) for the SOCS3 promoter region, including potential transcription factor binding sites (TFBS) Sp1, NFKB (diamonds) and within the CpG island (blue). (B) Percent DNA methylation at each individual CpG site investigated (18 CpG sites) in VAT of NGT (white; n = 22) vs. GDM group (GDM; black; n = 19). The overall mean across CpG sites is also included. Data are means ± S.E.M. TSS: Transcriptional start site, bp: base pairs.

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