Epigenetic Regulation of Myometrial Contractility in Pregnancy and Labor

妊娠和分娩过程中子宫肌收缩力的表观遗传调控

• 批准号: 10063452
• 负责人: CAROLE R MENDELSON
• 金额: $ 26.35万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-12-15 至 2023-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063452
• 关键词: Address; Aftercare; Anti-Inflammatory Agents; Binding; Biology; Birth; C-Terminal Binding Protein 1; Cells; Characteristics; Chromatin Structure; Collagen; Complex; DNA Binding Domain; DNA Polymerase II; Enzymes; Epigenetic Process; Estradiol; Euchromatin; Family; Family member; Gene Expression; Genes; Genetic; Genetic Transcription; Genomics; Goals; Heterochromatin; Histone H3; Hormones; Human; IL8 gene; Immunoblotting; Immunoprecipitation; Impairment; Induced Labor; Infant Mortality; Inflammatory; Knockout Mice; Life; Lipopolysaccharides; Mass Spectrum Analysis; Mediating; Metabolism; MicroRNAs; Mifepristone; Molecular; Mus; Myometrial; NF-kappa B; PTGS2 gene; Pathway interactions; Pregnancy; Pregnancy Maintenance; Premature Birth; Premature Labor; Progesterone; Progesterone Receptors; Protein Isoforms; Proteins; RNA; Regulation; Reporting; Research; Response Elements; Role; Time; Tissues; Transcription Factor AP-1; Transcription Repressor; Up-Regulation; Woman; base; chromatin immunoprecipitation; chromatin modification; deep sequencing; epigenetic regulation; gene discovery; genetic corepressor; histone modification; hormone regulation; insight; knock-down; member; mouse model; myometrium; next generation sequencing; novel; overexpression; p65; pregnant; prevent; progesterone receptor A; progesterone receptor B; promoter; receptor function; recruit; therapeutic target; transcription factor; transcriptome; transcriptome sequencing

PROJECT SUMMARY/ABSTRACT Preterm birth is the leading cause of infant mortality during the first four weeks of life world-wide. The overarching goal of this research is to enhance our understanding of the genetic and epigenetic mechanisms that mediate myometrial quiescence and contractility during pregnancy and labor, and their dysregulation leading to preterm birth. We suggest that progesterone (P4)/progesterone receptor (PR) maintains myometrial quiescence throughout most of pregnancy by inhibiting expression of inflammatory (e.g. IL-1b, IL-8, COX-2) and contractile (CAP) (e.g. OXTR, CX43) genes. We propose that this occurs via two basic mechanisms: (1) PR may tether to transcription factors (e.g. NF-kB, AP-1) bound to promoters of inflammatory genes and recruit corepressors to inhibit gene expression; (2) PR also may bind directly to promoters of genes encoding transcriptional repressors (e.g. ZEB1) to activate their expression. ZEB1, in turn, binds to promoters of CAP genes and recruits a repressive complex to inhibit their expression. By contrast, during the initiation of term and preterm labor, PR function in myometrium is impaired by its direct interaction with NF-kB, by increased expression of enzymes that metabolize P4 to inactive products, by decreased expression of PR coactivators and by upregulation of truncated PR isoforms (e.g. PR-A). The truncated PR isoforms may have reduced transcriptional and transrepressive activity. We postulate that these changes in inflammatory and CAP gene expression are mediated by alterations in chromatin modifications and structure. The goals of this proposal are to define the genes and mechanisms that underlie the inhibitory actions of P4/PR and differential actions of PR- A and PR-B on inflammatory and CAP gene expression, and to characterize the chromatin modifications that mediate myometrial quiescence and accompany enhanced CAP and inflammatory gene expression leading to term and preterm labor. To achieve these goals, we will use mouse models and human myometrial cells and tissues to: (1) define and characterize components of the complex of transcription factors/coregulators that interact with PR in the pregnant myometrium to mediate its anti-inflammatory actions and that interact with ZEB1 to inhibit CAP gene expression; (2) analyze expression and promoter-binding of these PR-interacting factors during pregnancy and with term and preterm labor and the effects of hormones and microRNAs in their regulation; (3) use RNA-seq and ChIP-seq to discover myometrial genes that are critical for the maintenance of pregnancy and initiation of labor and the underlying transcriptional mechanisms for their regulation. RNA-seq will enable discovery of genes involved in maintenance of pregnancy and the initiation of labor. The combined use of ChIP-seq will provide global insight into the transcriptional and epigenetic mechanisms that underlie these gene expression changes. Studies using PR-B-KO mice will elucidate the differential roles of PR-A vs. PR-B in regulation of the myometrial transcriptome. Collectively, our findings will reveal novel genes and pathways that can serve as therapeutic targets to prevent preterm birth and its consequences.

项目摘要/摘要 早产是全世界前四个星期的婴儿死亡率的主要原因。这 这项研究的总体目标是增强我们对遗传和表观遗传机制的理解 介导怀孕和劳动期间的子宫静脉静止和收缩力及其失调 导致早产。我们建议孕酮（P4）/孕激素受体（PR）保持子宫肌层 在整个怀孕的大部分时间里，静止通过抑制炎症的表达（例如IL-1B，IL-8，COX-2） 和收缩（CAP）（例如OXTR，CX43）基因。我们建议这是通过两种基本机制发生的：（1） PR可能与炎症基因启动子和 募集核压杆抑制基因表达； （2）PR还可以直接与编码基因的启动子结合 转录阻遏物（例如Zeb1）激活其表达。 Zeb1反过来与CAP启动子结合 基因和招募抑制性复合物以抑制其表达。相比之下，在开始期间 和早产，肌层中的PR功能受到与NF-KB的直接相互作用的损害 通过降低PR共激活剂的表达，将P4代谢为非活性产物的酶表达 并通过上调截短的PR同工型（例如PR-A）。截短的PR同工型可能减少了 转录和变性活动。我们假设这些炎症和CAP基因的变化 表达是由染色质修饰和结构的改变介导的。该提议的目标是 定义基因和机制，这些基因和机制是P4/PR的抑制作用以及PR的差异作用的基础 A和PR-B在炎症和CAP基因表达上，并表征染色质修饰 介导肌层静止，并伴随增强的帽和炎症基因表达，导致 术语和早产。为了实现这些目标，我们将使用鼠标模型和人肌层细胞，以及 组织至：（1）定义和表征转录因子/组合剂复合物的组成部分， 在怀孕的子宫肌层中与PR相互作用以介导其抗炎作用，并与之相互作用 ZEB1抑制CAP基因表达； （2）分析这些PR相互作用的表达和启动子结合 怀孕期间的因素，术语和早产以及激素和microRNA在其中的影响 规定; （3）使用RNA-SEQ和CHIP-SEQ发现对维持至关重要的子宫质基因 劳动的怀孕和启动及其调节的基本转录机制。 RNA-seq 将能够发现涉及维持妊娠和劳动启动的基因。组合 使用Chip-seq将提供全球洞察的洞察力，以了解基于的转录和表观遗传机制 这些基因表达会改变。使用PR-B-KO小鼠的研究将阐明PR-A V的差异作用。 PR-B在肌层转录组的调节中。总的来说，我们的发现将揭示新的基因和 可以作为预防早产及其后果的治疗靶标的途径。