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

描述(申请人提供)：每年全球将有330万婴儿死于导致早产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，感染约占美国结核病的20%-25%，是欠发达国家结核病的主要原因。宫颈重塑是分娩过程中的一个关键组成部分，在感染期间可能会被破坏。宫颈重塑是宫颈从封闭的僵硬结构转变为可以打开的结构，以便足月胎儿通过产道的过程。更好地了解足月和感染介导的早产宫颈重塑的机制将提供新的见解，可用于检测和预防肺结核。控制足月或早产宫颈重塑的过程在(1)转录水平上通过mRNAs、microRNAs和长非编码RNA(LncRNAs)的表达来调节，(2)在转录后水平上通过miRNAs对靶mRNAs和ncRNAs的作用来调节。这些机制的集成形成了一个调节电路，允许精细调整和仔细协调的基因表达程序。对完整转录组的鉴定，以及在相关生物学模型中microRNAs与其靶mRNAs和lncRNAs之间的临床相关相互作用，将为调节宫颈早熟的生物学机制提供新的见解。本研究的目标是应用最先进的基因组、生物信息学和计算方法来研究感染介导的早产小鼠模型中感染介导的基因调控，以及人类宫颈炎症的补充模型，以建立该方法的原则证明。具体地说，我们将使用RNA-SEQ和AGO HITS-CLIP(高通量RNAs测序)技术，结合基于细胞的基因特异性分析，来识别、确认和探索在这两种免费的宫颈感染/炎症模型中调控感染介导的宫颈成熟的基因表达程序。这项研究中确定的基因靶点的验证将支持这些方法未来的潜力，以剖析调控对足月成功分娩至关重要的过程的分子通路，并了解调控电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.