Gestational diabetes pathophysiology uncovered by placental transcriptomics

摘要 在怀孕期间，女性经历葡萄糖调节的主要生理变化，这些变化由以下因素调节： 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）。GDM是妊娠期最常见的代谢紊乱， 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。 用于预测GDM，并用于开发新的治疗剂以影响胰岛素分泌和胰岛素抵抗。 敏感性是妊娠期内和妊娠期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组研究， 测序和miRNA测序的尖端技术。胎盘样本（n=662）仔细 在一个名为Gen 3G的基于人群的孕妇前瞻性队列中收集。我们收集了 在整个妊娠期间，809名Gen 3G女性的精确表型和血浆样本，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，并客观地诊断 GDM。对于已鉴定的与体内胰岛素分泌相关的miRNA，我们将测试它们是否可以刺激胰岛素分泌， 在体外验证的β细胞模型中的胰岛素分泌。对于已鉴定的与胰岛素敏感性相关的miRNA， 将测试它们是否能影响体外脂肪细胞模型中的葡萄糖摄取。经过精心挑选， GDM与血糖正常妇女中最有希望的转录本，我们将开发检测方法， 妊娠早期收集的母体血浆中的分泌蛋白质。我们将测试新的 鉴定的胎盘特异性分泌蛋白（ELISA或质谱法）和可检测的miRNA（RT-qPCR） 妊娠早期血浆中GDM的发生率。我们将在两个前瞻性的多种族 怀孕妇女的队列。 该提案充分利用了独特的Gen 3G队列的丰富性，包括胎盘样本， 收集并保存用于转录组学研究、葡萄糖调节和血浆 整个妊娠期的样本。为这一申请而组建的调查小组具有丰富的专业知识 和以前的经验，以进行拟议研究的每一步。最后，我们的转录组结果将是 快速提供给更广泛的科学界的高度访问GTEx门户网站，因为我们将遵循他们所有的 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.