Defining Gene Expression Programs in Cervical Ripening: Roles for Non-Coding RNAs

描述（由申请人提供）：每年，全世界将有330万婴儿死于妊娠并发症，导致早产（PTB）或死产。虽然许多初始因素仍有待确定，但我们知道，在美国，感染约占PTB的20-25%，是欠发达国家PTB的主要原因。宫颈重塑是指宫颈从一个封闭的刚性结构转变为一个可以打开的结构，以允许足月胎儿通过产道的过程，这是分娩过程的关键组成部分，可能在感染期间被破坏。更好地了解驱动期和感染介导的早产颈椎重塑的机制将为PTB的检测和预防提供新的见解。足月或早产时颈椎重构的调控过程在(1)转录水平上通过mrna、microrna和长链非编码rna （lncrna）的表达进行调控；(2)转录后水平上通过mirna对靶mrna和ncrna的作用进行调控。这些机制的整合形成了一个调控回路，使基因表达程序得以微调和精心协调。完整转录组的鉴定，以及相关生物学模型中microrna与其靶mrna和lncrna之间临床相关的相互作用，将为介导宫颈早熟的生物学机制提供新的见解。当前研究的目标是应用尖端的基因组学、生物信息学和计算方法来研究感染介导的基因调控在感染介导早产的小鼠模型中，以及在人类子宫颈炎症的补充模型中，以建立该方法的原理证明。具体来说，我们将使用RNA-seq和Ago HITS-CLIP（通过Argonaute交联免疫沉淀分离的rna的高通量测序）技术，结合基于细胞的基因特异性测定，识别、确认和探索在这两种互补的宫颈感染/炎症模型中调节感染介导的宫颈成熟的基因表达程序。本研究中确定的基因靶点的验证将支持这些方法的未来潜力，以剖析对足月成功分娩至关重要的调节过程的分子途径，并了解调节回路如何在早产中出错。

DESCRIPTION (provided by applicant): On an annual basis, 3.3 million babies will die worldwide due to complications in pregnancy that leads to preterm birth (PTB) or stillbirths. While many of the initiating factors remain to be identified, we know that infection accounts for roughly 20-25% of PTBs in the United States and is the primary cause of PTB in underdeveloped countries. Cervical remodeling - the process by which the cervix is transformed from a closed rigid structure to one that can open to allow passage of a term fetus through the birth canal - is a key component of the birth process that may be disrupted during infections. A better understanding of mechanisms that drive term and infection-mediated preterm cervical remodeling will provide new insights that can be used for the detection and prevention of PTB. The processes that govern cervical remodeling in term or preterm birth are regulated at (1) the transcriptional level by the expression of mRNAs, microRNAs, and long non-coding RNAs (lncRNAs) and (2) the post-transcriptional level by the actions of miRNAs on target mRNAs and ncRNAs. The integration of these mechanisms forms a regulatory circuit that allows finely tuned and carefully coordinated gene expression programs. The identification of the complete transcriptome, as well as clinically relevant interactions between microRNAs and their target mRNAs and lncRNAs in relevant biological models, will provide new insights into the biological mechanisms that mediate premature cervical ripening. The goal of the current study is to apply cutting-edge genomic, bioinformatic, and computational approaches to the study of infection-mediated gene regulation in a mouse model of infection mediated preterm birth, as well as a complementary model of inflammation in the human cervix to establish proof-of-principle for this approach. Specifically, we will use RNA-seq and Ago HITS-CLIP (high-throughput sequencing of RNAs isolated by crosslinking immunoprecipitation of Argonaute) technologies, in conjunction with cell-based gene-specific assays, to identify, confirm, and explore the gene expression programs that regulate infection mediated cervical ripening in these two complimentary models of cervical infection/inflammation. Validation of gene targets identified in this study will suppor the future potential of these approaches to dissect the molecular pathways that regulate processes critical for successful parturition at term and to understand how regulatory circuits go awry in preterm birth.