The Role of Corticotrophin Releasing Hormone on Placental Transcriptional Networks and Birth Timing

项目摘要 我的长期职业目标是建立一个独立的研究线使用数据驱动， 综合方法来检查分娩，我的兴趣在于糖皮质激素信号 怀孕期间促肾上腺皮质激素释放激素（CRH）是一种肽类激素， 糖皮质激素信号也由胎盘产生。CRH影响胎盘生长 和功能，调节母体代谢，对胎儿发育至关重要。胎盘CRH 在妊娠过程中释放到母体血浆中，在分娩前达到峰值， 妊娠中期CRH升高预示早产。这个问题的核心假设是 有人认为，妊娠中期的CRH水平改变了胎盘基因表达的格局， 对胎盘功能和妊娠期长短的下游影响。这种多学科 项目将汇集共同导师谁是在各自领域的关键专家;博士内森 Price（系统生物学）、Sheela Sathyanarayana博士（流行病学）和Louis Muglia博士 （怀孕的内分泌调节）。通过Price实验室开发的软件， 产生了一个大规模的人类胎盘转录调控网络（TRN）。在这 建议的分析，我们将扩大这个TRN，以确定相关的共享转录驱动程序， 母亲血浆CRH和妊娠期长度，使用的样本来自于 华盛顿回声大道。在目标1中，我们将产生一个全基因组的，胎盘特异性的， 建立了转录因子与其靶基因之间的定量关系模型，并验证了 该模型使用CRISPR/Cas9技术编辑关键转录因子并量化变化， 在下游基因表达中。在目标2中，我们将鉴定与CRH相关的基因， 以及与孕龄相关的基因，然后确定重叠基因， 丰富了与这些基因相关的生物学途径。在目标3中，我们将利用TRN来识别 TF模块（基因网络，其表达通过共同的 转录因子），其与目的2中鉴定的差异表达基因相关， 我们将通过胎盘来源的细胞系进行体外验证。这样，我们就能识别 与母体血浆CRH和妊娠时间相关的共同转录模块。我们 预计这项数据驱动的研究将揭示涉及CRH在以下方面作用的分子机制： 间隔和妊娠期长度，并提供潜在的目标，以延长妊娠期长度， 妊娠并发早产。此外，这项工作将为我提供 建立独立研究线所需的背景。

PROJECT SUMMARY My long-term career objective is to establish an independent line of research using data driven, integrated approaches to examine parturition, and my interests lie within glucocorticoid signaling in pregnancy. Corticotrophin releasing hormone (CRH) is a peptide hormone involved in glucocorticoid signaling that is also produced by the placenta. CRH influences placental growth and function, modulates maternal metabolism, and is crucial for fetal development. Placental CRH is released into maternal plasma over the course of gestation, peaking before parturition, and elevated CRH in mid gestation is predictive of preterm birth. The central hypothesis of this proposal is that CRH levels in mid pregnancy alter the placental gene expression landscape, with downstream consequences on placental function and gestational length. This multidisciplinary project will bring together co-mentors who are key experts in their respective fields; Dr. Nathan Price (systems biology), Dr. Sheela Sathyanarayana (epidemiology) and Dr. Louis Muglia (endocrine regulation of pregnancy). Through software developed within in the Price lab, we have generated a large scale transcriptional regulatory network (TRN) of the human placenta. In this proposed analysis, we will expand this TRN to identify shared transcriptional drivers related to both maternal plasma CRH and gestational length, using samples from the University of Washington ECHO PATHWAYS. In aim 1, we will generate a genome wide, placental specific model quantifying relationships between transcription factors and their target genes, and validate this model using CRISPR/Cas9 technology to edit key transcription factors and quantify changes in downstream gene expression. In aim 2, we will identify genes related to CRH measured in mid to late gestation as well as genes related to gestational age, then identify overlapping genes and enriched biological pathways related to these genes. In aim 3, we will leverage the TRN to identify TF modules (networks of genes whose expression coordinately changes through a common transcription factor) which are related to the differentially expressed genes identified in aim 2, which we will validate in vitro through placental derived cell lines. In this way, we will identify shared transcriptional modules related to maternal plasma CRH and gestational length. We anticipate that this data driven study will uncover molecular mechanisms involving CRH's role in partition and gestational length, and provide potential targets to lengthen gestational length in pregnancies complicated by preterm birth. Additionally, this work will provide me with the background needed to establish an independent line of research.