Mechanisms of infection-mediated cervical ripening

 描述（由申请人提供）：在美国，感染占早产的25-40%，是不发达国家早产的主要原因。有证据表明，分娩的准备工作在妊娠早期就开始了，早产的机制与足月不同，并取决于病因，强调需要定义特定的通路调控机制。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，在足月和早产中子宫收缩开始之前。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。在相关生物学模型中鉴定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。本研究的目标是通过计算查询最近生成的宫颈polyA+ RNA-Seq和microRNA微阵列数据集，这些数据集来自足月和感染介导的早产小鼠模型，以识别受调控的microRNA及其预测的mRNA和lncRNA靶标。将建立3D-人宫颈基质细胞培养系统和基于细胞的基因特异性测定，以验证将使用特定标准选择的~ 10种microRNA：靶RNA相互作用。最后，感染介导的早产对产后宫颈修复和宫颈功能/第二次妊娠早产风险的影响将使用组织生物力学，生理学以及尖端成像和基因组方法进行全面研究。总的来说，这些研究将剖析调节足月成功分娩关键过程的分子途径，并定义感染介导的早产中出错的调节回路。

 DESCRIPTION (provided by applicant): Infection accounts for 25-40% preterm births in the United States and is the primary cause of preterm birth in underdeveloped countries. Evidence that preparation for parturition begins early in pregnancy and that mechanisms of preterm birth are distinct from term and dependent on etiology, emphasize the need to define pathway specific regulatory mechanisms. 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 precedes onset of uterine contractions in term and preterm birth. 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 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 computationally interrogate recently generated cervical polyA+ RNA-Seq and microRNA microarray datasets from term and infection-mediated preterm mouse models to identify regulated microRNAs and their predicted mRNAs and lncRNA targets. A 3D-human cervical stromal cell culture system and cell based gene specific assays will be established in order to validate ~ 10 microRNA:target RNA interactions that will be selected using specific criteria such. Finally the impact of an infection-mediated preterm birth on postpartum cervical repair and cervical function/risk of prematurity in a second pregnancy will be investigated comprehensively using tissue biomechanics, physiology as well as cutting edge imaging and genomic approaches. Collectively these studies will dissect the molecular pathways that regulate processes critical for successful parturition at term and define the regulatory circuits that go awry in infection-mediated preterm birth.