Gestational diabetes pathophysiology uncovered by placental transcriptomics

摘要摘要 在怀孕期间，女性经历了葡萄糖调节的主要生理变化，这些变化是由 胎盘。在一些孕妇中，胰岛素敏感性和分泌之间的调节是不平衡的， 导致妊娠期糖尿病(GDM)。妊娠期糖尿病是妊娠期最常见的代谢紊乱 会导致母亲和后代在怀孕期间和怀孕后很长时间内出现并发症。更好 了解胎盘在妊娠期血糖调节中的作用对于识别早期生物标志物是必要的 预测妊娠期糖尿病，并开发新的治疗药物来影响胰岛素分泌和胰岛素 敏感性，孕期内外糖尿病的两个主要病理生理决定因素。 我们建议使用最新的RNA方法进行胎盘全基因组转录研究- 测序和尖端的miRNA测序技术。胎盘标本(n=662)仔细检查 这是在一个名为Gen3G的孕妇群体预期队列中收集的。我们收集了 809名第三代女性妊娠期的精制表型和血浆样本，包括口服葡萄糖 耐受性测试使我们能够表征他们的胰岛素分泌、胰岛素敏感性和客观诊断 GDM。对于已确定的与体内胰岛素分泌相关的miRNA，我们将测试它们是否能够刺激 体外验证的β细胞模型中的胰岛素分泌。对于已确定的与胰岛素敏感性相关的miRNA，我们 将在体外脂肪细胞模型中测试它们是否会影响葡萄糖的摄取。经过精心挑选 最有希望的GDM和正常血糖女性的转录本，我们将开发检测方法来检测 妊娠早期采集的母体血浆中的分泌蛋白。我们将测试新的 可检测到的胎盘特异性分泌蛋白(EL ISA或MS)和miRNA(RT-qPCR) 妊娠早期血浆中GDM的发生率。我们将在两个预期的多种族人群中复制我们的发现 一群怀孕的妇女。 这项建议利用了独特的Gen3G队列的丰富性，包括仔细的胎盘样本 收集和保存以用于转录研究、精炼的血糖调节表型和血浆 怀孕期间的样本。为这一应用程序组建的调查团队拥有丰富的专业知识 和以前的经验来进行拟议研究的每一步。最后，我们的转录结果将是 在高度访问的GTEx门户网站上迅速提供给更广泛的科学界，因为我们将跟踪他们的所有 我们测序的标准程序。

Summary abstract During pregnancy, women experience major physiologic changes in glucose regulation that are regulated by the placenta. In some pregnant women, regulation between insulin sensitivity and secretion is imbalanced, leading to gestational diabetes mellitus (GDM). GDM is the most common metabolic disorder in pregnancy and leads to complications in both mothers and offspring during pregnancy and long afterwards. Better understanding of the placental role in gestational glucose regulation is necessary to identify early biomarkers that predict GDM, and for the development of novel therapeutic agents to influence insulin secretion and insulin sensitivity, the two major pathophysiologic determinants of diabetes inside and outside of pregnancy.. We propose to conduct placental genome-wide transcriptomic studies using the most recent methods for RNA- sequencing and cutting-edge technology for miRNA sequencing. Placenta samples (n=662) were carefully collected in a population-based prospective cohort of pregnant women called Gen3G. We have collected refined phenotypes and plasma samples in 809 Gen3G women across gestation, including oral glucose tolerance tests allowing us to characterize their insulin secretion, insulin sensitivity, and objectively diagnosed GDM. For identified miRNA associated with insulin secretion in vivo, we will test whether they can stimulate insulin secretion in in vitro validated β-cell models. For identified miRNA associated with insulin sensitivity, we will test whether they can influence glucose-uptake in in vitro adipocytes models. After meticulous selection of the most promising transcripts from our GDM vs normoglycemic women, we will develop assays to detect secreted proteins in maternal plasma collected at 1st trimester. We will test associations between newly identified placenta-specific secreted proteins (ELISA or Mass Spectrometry) and miRNA (RT-qPCR) detectable in plasma at 1st trimester with GDM incidence. We will replicate our findings in two prospective multi-ethnic cohorts of pregnant women. This proposal leverages the richness of the unique Gen3G cohort, including placenta samples carefully collected and preserved for transcriptomic studies, refined phenotypes of glucose regulation and plasma samples across gestation. The team of investigators assembled for this application has substantial expertise and previous experience to conduct each step of the proposed study. Finally, our transcriptomic results will be rapidly available to the wider scientific community on the highly accessed GTEx portal, as we will follow all their standard procedures for our sequencing.