Human Feto-placental Arterial and Venous Endothelial Cells are Differentially Programmed by Gestational Diabetes Mellitus Resulting in Cell-specific Barrier Function Changes. Homo sapiens

We performed genome-wide methylation analysis of primary feto-placental arterial and venous endothelial cells from healthy (AEC and VEC) and GDM complicated pregnancies (dAEC and dVEC). Parallel transcriptome analysis identified variation in gene expression linked to GDM-associated DNA methylation, implying a direct functional link. Pathway analysis found that genes altered by exposure to GDM clustered to functions associated with ’Cell Morphology’ and ’Cellular Movement’ in both AEC and VEC. Further functional analysis demonstrated that GDM exposed cells have altered actin organization and barrier function. Our data indicate that exposure to GDM programs atypical morphology and barrier function in feto-placental endothelial cells by DNA methylation and gene expression change. The effects differ between AEC and VEC, indicating a stringent cell-specific sensitivity to adverse exposures associated with developmental programming in utero. Overall design: Primary human feto-placental endothelial cells form arteries (AEC) and veins (VEC) were isolated after healthy pregnancy (AEC, VEC) and after a pregnancy complicated by Gestational Diabetes Mellitus (dAEC, dVEC). Total RNA was isolated and subjected to microarray analysis.

本研究对健康妊娠（动脉内皮细胞（arterial endothelial cells, AEC）与静脉内皮细胞（venous endothelial cells, VEC））及妊娠期糖尿病（Gestational Diabetes Mellitus, GDM）合并妊娠（GDM暴露动脉内皮细胞（dAEC）与GDM暴露静脉内皮细胞（dVEC））来源的原代胎儿胎盘动脉内皮细胞与静脉内皮细胞，开展了全基因组甲基化分析。平行转录组分析发现，与GDM相关的DNA甲基化伴随基因表达变异，提示二者存在直接的功能关联。通路富集分析显示，在AEC与VEC中，受GDM暴露影响的差异基因均富集于"细胞形态"与"细胞运动"相关功能通路。进一步功能实验证实，经GDM暴露的细胞其肌动蛋白组织与屏障功能均发生异常改变。本研究数据表明，GDM暴露可通过DNA甲基化与基因表达调控，使胎儿胎盘内皮细胞出现非典型形态与屏障功能异常。且该效应在AEC与VEC间存在显著差异，提示胎儿胎盘内皮细胞对宫内发育程序化相关的不良暴露具有严格的细胞特异性敏感性。整体实验设计：分别从健康妊娠及GDM合并妊娠的胎盘中，分离得到动脉来源（AEC）与静脉来源（VEC）的原代人胎儿胎盘内皮细胞，其中健康妊娠组的细胞记为AEC、VEC，GDM合并妊娠组的细胞记为dAEC、dVEC。提取总RNA后进行基因芯片（microarray）分析。