Metformin Effects Placental Trophoblast Function

项目摘要 最佳的胎儿生长和发育需要强大的胎盘功能，这是受血糖和 脂质浓度以及潜在的化学品/药物暴露。药物可能会通过以下方式改变表观遗传稳态 直接或间接的机制（转录活性），可以在暴露于药物后长期存在（即， 药物表观基因组学）。我们的初步数据显示，在DNA甲基化的差异区域， 与非GDM滋养细胞相比，妊娠合并妊娠糖尿病（GDM）的滋养细胞。 此外，二甲双胍和GDM独立且显着降低线粒体呼吸， 胎盘滋养层细胞中胎盘激素基因和蛋白的表达。胎盘激素，尤其是β- hCG是胎儿正常生长和发育所必需。最近在妊娠期使用二甲双胍的试验 显示出潜在的母体益处，如较低的妊娠期体重增加，但较高的小胎儿发生率， 胎龄婴儿和更大的体重z分数和肥胖的儿童。这突出表明， 我们对二甲双胍对胎盘健康的影响及其在潜在的有益和不利影响方面的作用的了解 围产期结果。我们的目标是（目的1）确定二甲双胍对滋养层细胞表观基因组的影响， 和转录组学特征，并（目的2）确定二甲双胍治疗抑制 GDM和非GDM胎盘中细胞滋养层成熟和hCG产生。表观基因组和 来自培养8小时（细胞滋养层）和72小时的分离的滋养层的转录组谱 将暴露于二甲双胍的合体滋养细胞与来自GDM和非合体滋养细胞的未暴露滋养细胞进行比较。 GDM妊娠活细胞，整个滋养层成熟过程中的时间推移成像（体外8、24、48、72小时 将进行形态学成熟与滋养层的基因标志物的视觉关联 在存在或不存在二甲双胍细胞暴露的情况下，分化和β-hCG产生。二甲双胍和 GDM必须同时研究，因为已发表的试验数据（即肥胖和多囊卵巢综合征） 综合征）已经表明它们与胎儿的生长和健康独立相关。我们 我承认二甲双胍已经在许多器官系统中进行了研究，但它如何影响胎盘， 对滋养层细胞的了解很少。我们认为，重要的是要了解， 二甲双胍影响胎盘滋养层细胞的成熟和代谢以及表观遗传机制 在GDM和非GDM中调节滋养层功能的基因表达模式的变化 胎盘这项研究将提供新的信息，可能会警告在怀孕期间使用二甲双胍，并设定 进行更精细的临床试验。

PROJECT SUMMARY Optimal fetal growth and development requires robust placental function, which is affected by blood glucose and lipid concentrations but also potentially chemical/drug exposures. Drugs may alter epigenetic homeostasis by direct or indirect mechanisms (transcription activity) that can persist long after exposure to the drug (i.e. pharmacoepigenomics). Our preliminary data has shown differentially methylated regions in DNA from trophoblasts in pregnancies complicated by gestational diabetes (GDM) compared to non-GDM trophoblasts. Moreover, metformin and GDM independently and significantly decreases mitochondrial respiration and placental hormone gene and protein expression in placental trophoblasts. Placental hormones, particularly β- hCG are necessary for normal fetal growth and development. Recent trials using metformin in pregnancy has demonstrated potential maternal benefits such as lower gestational weight gain but higher rates of small-for- gestational age infants and greater weight z-scores and adiposity in children. This highlights the critical gap in our knowledge of the effects of metformin on placental health and its role on potential beneficial and adverse perinatal outcomes. Our objectives are to (Aim 1) determine the effects of metformin on trophoblast epigenomic and transcriptomic profiles and (Aim 2) determine the degree to which metformin treatment suppresses cytotrophoblast maturation and hCG production in GDM and non-GDM placentas. Epigenomic and transcriptomic profiles from isolated trophoblasts in culture for 8-hrs (cytotrophoblasts) and 72-hrs (syncytiotrophoblasts) exposed to metformin will be compared to unexposed trophoblasts from GDM and non- GDM pregnancies. Live cell, time-lapsed imaging throughout trophoblast maturation (8, 24, 48, 72-hrs in vitro culture) will be done to visually correlate morphological maturation with gene markers of trophoblast differentiation and β-hCG production in the presence or absence of metformin cell exposure. Metformin and GDM must be studied together and concurrently because published trial data (i.e. obesity and polycystic ovarian syndrome) has shown they are independently associated with the growth and health of the fetus. We acknowledge metformin has been studied in many organ systems but how it effects the placenta and specifically the trophoblast cells is poorly understood. We believe that it is important to understand the degree to which metformin affects the maturation and metabolism of placental trophoblasts and whether epigenetic mechanisms underlie changes in gene expression patterns that regulate trophoblast function in GDM and non-GDM placentas. This study will offer new information that may caution the use of metformin in pregnancy and set the stage for more refined clinical trials.