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

描述（由申请人提供）：每年，全球将有330万婴儿死于导致早产（PTB）或死产的妊娠并发症。虽然许多引发因素仍有待确定，但我们知道，感染约占美国PTB的20-25%，是不发达国家PTB的主要原因。子宫颈重塑-子宫颈从封闭的刚性结构转变为可以打开以允许足月胎儿通过产道的过程-是分娩过程的关键组成部分，可能在感染期间被破坏。更好地了解驱动足月和感染介导的早产儿宫颈重塑的机制将提供可用于检测和预防PTB的新见解。在足月或早产中控制宫颈重塑的过程在（1）转录水平通过mRNA、microRNA和长链非编码RNA（lncRNA）的表达来调节，以及（2）转录后水平通过miRNA对靶mRNA和ncRNA的作用来调节。这些机制的整合形成了一个调控回路，允许精细调节和仔细协调的基因表达程序。完整转录组的鉴定，以及相关生物模型中microRNA与其靶mRNA和lncRNA之间的临床相关相互作用，将为介导宫颈过早成熟的生物学机制提供新的见解。目前研究的目标是将尖端的基因组学，生物信息学和计算方法应用于感染介导的早产小鼠模型中感染介导的基因调控的研究，以及人类宫颈炎症的补充模型，以建立这种方法的原理证明。具体来说，我们将使用RNA-seq和Ago HITS-CLIP（通过Argonaute的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，以识别，确认和探索在这两种宫颈感染/炎症的互补模型中调节感染介导的宫颈成熟的基因表达程序。在这项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，并了解早产中调节回路如何出错。

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.