Influence of gestational diabetes mellitus on human umbilical vein endothelial cell miRNA

Abstract

We aimed to identify miRNAs whose expression levels in fetal tissues are altered by exposure to a diabetic milieu and elucidate the impact on target protein expression. Gestational diabetes mellitus (GDM) affects both immediate and future disease risk in the offspring. We hypothesized that GDM alters miRNA expression in human umbilical vein endothelial cells (HUVECs) that may influence metabolic processes. A cross-sectional design compared differences in miRNA expression in HUVECs and target protein abundance in placentae between infants of women with GDM (IGDM) and infants born to normoglycaemic controls. miRNAs were identified using microarray profiling and literature review and validated by quantitative PCR (qPCR). In vitro transfection studies explored the impact of the miRNA on target protein expression. Expression of seven miRNA species, miR-30c-5p, miR-452-5p, miR-126-3p, miR-130b-3p, miR-148a-3p, miR-let-7a-5p and miR-let-7g-5p, was higher in the HUVECs of IGDM. Abundance of the catalytic subunit of AMP-activated protein kinase α1 (AMPKα1) was decreased in the HUVECs and BeWo cells (transformed trophoblast cell line) transfected with miR-130b and miR-148a mimics. AMPKα1 expression was also decreased in placental tissues of IGDM. The expression of several miRNAs were altered by in utero exposure to DM in infants of women whose dysglycaemia was very well controlled by current standards. Decreased expression of AMPKα1 as a result of increased levels of miR-130b and miR-148a may potentially explain the decrease in fat oxidation we reported in infants at 1 month of age and, if persistent, may predispose offspring to future metabolic disease.

Role of microRNA-130b in placental PGC-1α/TFAM mitochondrial biogenesis pathway

“…Further studies examining expression of miR-130b-3p and the PGC-1α/TFAM pathway in intact placentae and or primary cultured trophoblasts are warranted. In addition to PGC-1α, miR-130b-3p is known to target other proteins involved in energy metabolism, such as AMP-activated protein kinase α1 (AMPKα1) [20, 27]. We recently reported that placental AMPKα1 is also reduced in diabetic women [20].…”

“…In addition to PGC-1α, miR-130b-3p is known to target other proteins involved in energy metabolism, such as AMP-activated protein kinase α1 (AMPKα1) [20, 27]. We recently reported that placental AMPKα1 is also reduced in diabetic women [20]. Thus miR-130b-3p could be acting indirectly or additively through AMPK, since activated AMPK can upregulate PGC-1α expression [32].…”

“…All qPCR reactions for miRNAs were performed using TaqMan Universal Master Mix II, no UNG (Life Technologies, Grand Island, NY) as previously described [20]. Relative expression of individual miRNA species was calculated using the 2 −ΔΔC T method [21] with the software DataAssist, where RNU48 served as the endogenous control miRNA.…”

“…These small non-coding RNAs negatively regulate abundance of proteins involved in various physiological and pathological cellular processes by affecting translation or stability of targeted mRNAs [19]. Previous studies in our laboratory demonstrated increased expression of several microRNAs, including microRNA-130b (miR-130b-3p) and microRNA-148a (miR-148a-3p), in human umbilical vein endothelial cells from offspring of diabetic mothers [20]. These 2 miRNAs in particular has been predicted to target PGC-1α.…”

Effects of maternal diabetes and fetal sex on human placenta mitochondrial biogenesis

“…Our previous studies demonstrated a decrease in the catalytic subunits of AMP activated protein kinase (AMPKα1) in placentae of diabetic women [23]. AMPK plays an essential role in maintaining cell energy homeostasis and it regulates the expression and activation of PGC-1α, the master regulator of mitochondrial biogenesis and energy metabolism [24–26].…”