The role of Corticotrophin Releasing Hormone on placental transcriptional networks and birth timing

项目概要 我的长期职业目标是利用数据驱动建立独立的研究路线， 检查分娩的综合方法，我的兴趣在于糖皮质激素信号传导 在怀孕期间。促肾上腺皮质激素释放激素 (CRH) 是一种肽激素，参与 糖皮质激素信号也由胎盘产生。 CRH影响胎盘生长 和功能，调节母体新陈代谢，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，并且 妊娠中期 CRH 升高可预测早产。这个假设的中心假设 建议认为妊娠中期的 CRH 水平会改变胎盘基因表达景观， 对胎盘功能和妊娠长度的下游影响。这个多学科 项目将汇集各自领域的关键专家共同导师；内森博士 Price（系统生物学）、Sheela Sathyanarayana 博士（流行病学）和 Louis Muglia 博士 （妊娠内分泌调节）。通过 Price 实验室开发的软件，我们 生成了人类胎盘的大规模转录调控网络（TRN）。在这个 提出的分析，我们将扩展此 TRN 以识别与以下内容相关的共享转录驱动程序 母体血浆 CRH 和妊娠长度，使用来自大学的样本 华盛顿回声通路。在目标 1 中，我们将生成一个全基因组、胎盘特异性的 建立转录因子与其靶基因之间的量化关系模型，并验证 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，我们将鉴定与中期测量的 CRH 相关的基因。 到妊娠晚期以及与胎龄相关的基因，然后识别重叠的基因和 丰富了与这些基因相关的生物途径。在目标 3 中，我们将利用 TRN 来识别 TF 模块（基因网络，其表达通过共同的协调变化 转录因子），与目标 2 中确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。这样我们就可以识别出 与母体血浆 CRH 和妊娠长度相关的共享转录模块。我们 预计这项数据驱动的研究将揭示涉及 CRH 作用的分子机制 分区和妊娠长度，并提供延长妊娠长度的潜在目标 妊娠并发早产。此外，这项工作将为我提供 建立独立研究路线所需的背景。

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.