Biological Roles of Poly(ADP-ribose) Polymerase-1 in Preterm Labor

早产是影响全世界家庭的最常见的妊娠并发症，也是发达国家围产期发病率和死亡率的主要原因。长期的研究目标是确定在足月和早产时调节分娩的关键生理和遗传途径，以治疗和预防肺结核。在分娩过程中，子宫肌层高度收缩和兴奋，从而对炎症信号和前列腺素做出反应，产生协调而有力的分娩收缩。值得注意的是，感染和感染引发的炎症可能导致25%-40%的早产。特发性早产和感染介导的早产的一个共同因素是炎症状态的存在。不同细胞类型的炎症反应共享一组共同的、协调良好的分子事件，导致促炎细胞因子和其他促炎蛋白的表达。在许多疾病状态下，广泛的细胞应激触发不受控制的聚(ADP-核糖)聚合酶-1(PARP-1)激活。早期的研究表明，PARP-1缺失的小鼠对感染性休克具有抵抗力，最近的研究强调了PARP-1在应对炎症应激信号中的作用。本研究主要研究PARP-1在调节子宫肌层对病理性炎症和收缩反应中的作用。在强大的初步数据和使用PARP抑制剂(PARPI)治疗炎症性疾病的良好结果的指导下，提出了三个特定的目标：(1)确定PARP-1在PTB中的活性程度，并探索PARPI作为一种潜在的延缓分娩开始的治疗方法；(2)确定PARP-1对子宫生理的影响；以及(3)确定PARP-1在介导转录因子功能中的作用。综合运用小鼠模型、基因组学和蛋白质组学的方法来阐明PARP-1依赖的调节子宫肌层生理学和PTB的机制的作用。拟议的目标在概念和技术上都是创新的，共同填补了我们在子宫生物学和肺结核基础知识方面的重大空白，将对该领域产生广泛的影响。此外，目前的提议将使我能够获得以下方面的技术专长：(1)最先进的基因组方法(PRO-SEQ和CHIP-SEQ)，(2)研究PALP的新方法(ADPR检测试剂，asPARP质谱仪)和(3)先进的计算方法。此外，克劳斯实验室和德克萨斯大学西南医学中心提供了一个环境，将促进我作为一名科学家的成长，并成功过渡到一名独立的调查员

Premature birth (PTB) is the most common complication of pregnancy affecting families worldwide and is the leading cause of perinatal morbidity and mortality in developed countries. The long-term research goal is to define critical physiological and genetic pathways that regulate labor at term and preterm in order to treat and prevent PTB. During the parturition process, the myometrium becomes highly contractile and excitable to produce the coordinated and forceful contractions of labor in response to inflammatory signals and prostaglandins. Of note, infection and infection-driven inflammation may contribute to 25–40% of preterm births. A common element of both idiopathic and infection-mediated preterm is the presence of an inflammatory state. Inflammatory responses in different cell types share a common set of well-coordinated molecular events that lead to the expression of proinflammatory cytokines and other proinflammatory proteins. In many disease states, extensive cellular stress triggers uncontrolled poly(ADP-ribose) polymerase-1 (PARP-1) activation. Early studies showed that PARP-1 null mice are resistant to septic shock and more recent studies have highlighted the roles of PARP-1 in response to inflammatory stress signals. This application is specifically focused on characterizing the role of PARP-1 in mediating the myometrial response to pathological (LPS)-mediated inflammation and contractility. Guided by strong preliminary data and promising results using PARP inhibitors (PARPi) to treat inflammatory disease, three specific aims are proposed: (1) determine the extent of PARP-1 activity in PTB and explore PARPi as a potential therapeutic in delaying the onset of labor; (2) determine the impact of PARP-1 on uterine physiology in response to LPS-mediated PTB; and (3) determine the role of PARP-1 in mediating transcription factor function. An integrative approach employing a combination of mouse models, genomics, and proteomics will be used to elucidate the role of PARP-1-dependent mechanisms regulating myometrial physiology and PTB. The proposed aims are conceptually and technically innovative and together will have a broad impact on the field by filling a substantial gap in our fundamental knowledge of uterine biology and PTB. In addition, the current proposal will allow me to gain technical expertise in (1) state-of-the-art genomic methodologies (PRO-seq and ChIP-seq), (2) novel methods to study PARPs (ADPR-detection reagents, asPARP mass spec) and (3) advanced computational methods. Further, the Kraus lab and UT Southwestern Medical Center provides an environment that will facilitate my growth as a scientist and successful transition to becoming an independent investigator