Static state of epithelial mesenchymal transition in fetal membrane cells: a novel inflammatory pathway to parturition

胎儿膜细胞上皮间质转化的静态：一种新的分娩炎症途径

• 批准号: 9893012
• 负责人: RAMKUMAR MENON
• 金额: $ 7.9万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9893012
• 关键词: Aging; Binding Proteins; Biological; Biological Assay; Birth; Cell Culture Techniques; Cell membrane; Cells; Characteristics; Complement Factor B; Cytokeratin 18; Data; E-Cadherin; Embryo; Endocrine; Environment; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Epithelium; Equilibrium; Exposure to; Fetal Growth; Fetal Membranes; Fetal Tissues; Fetus; Gene Silencing; Growth Factor; Homeostasis; Hormones; Immune; In Vitro; Induced Labor; Infection; Inflammation; Inflammatory; Inflammatory Response; Injury; Intervention; MAP3K7IP1 gene; MMP9 gene; Maintenance; Mechanics; Mediating; Membrane; Mesenchymal; Methods; Modeling; Molecular; N-Cadherin; Oxidative Stress; Pathologic; Pathway interactions; Phosphotransferases; Physiological; Play; Pregnancy; Premature Birth; Premature Labor; Prevention; Process; Production; Progesterone; RNA Interference; Reporting; Role; Signal Pathway; Signal Transduction; Site; Small Interfering RNA; Structure; TWIST1 gene; Testing; Tissues; Transforming Growth Factor beta; Transforming Growth Factor beta Receptors; Transforming Growth Factors; Uterus; Vimentin; Work; adverse pregnancy outcome; amnion; cell motility; epithelial to mesenchymal transition; experimental study; in utero; inhibitor/antagonist; injured; novel; p38 Mitogen Activated Protein Kinase; premature; preterm premature rupture of membranes; prevent; receptor; repaired; response; senescence; transcription factor; wound healing

ABSTRACT Fetal membranes (amniochorion) provide the structural framework and perform mechanical and protective functions during pregnancy. A progressive p38 mitogen-activated protein kinase (MAPK)-mediated senescence (mechanism of aging) occurs in fetal membranes. It is correlated with fetal growth and is accelerated at term due to increased oxidative stress (OS) in the intrauterine cavity. Senescence of fetal tissues causes inflammation that can promote labor; thus, membranes play a critical role in pregnancy and parturition. Throughout gestation, membranes maintain homeostasis by repairing themselves when cells are shed and the matrix is degraded. This remodeling process creates gaps and microfractures. At the cellular level, it remains unclear how the membranes' pluripotent amnion epithelial and mesenchymal cells repair the damaged sites. Since remodeling is essential for maintaining membrane homeostasis and preventing adverse pregnancy outcomes, understanding this process is critical. We have determined that injury to amnion epithelial cells (AEC) forces AEC proliferation and cell migration to rebuild injured sites, which is a normal physiologic response. Pilot data suggest that OS- and p38 MAPK-mediated senescence cause epithelial mesenchymal transition (EMT) and prevent tissue remodeling through increased production of TGFβ. We also determined that TGFβ plays dual functions: 1) induction of senescence by autophosphorylation of p38MAPK and 2) induction of EMT transcription factors to facilitate cellular and mechanical membrane disruption. These effects were reversed by progesterone (P4). We hypothesize that balanced tissue remodeling maintains fetal membrane homeostasis during pregnancy, but TGFβ increase due to overwhelming OS at term or in response to infection and inflammation (TNFα) at preterm causes an irreversible state of p38MAPK-mediated senescence and EMT that can promote membrane damage. Additionally, we hypothesize that during pregnancy, TGFβ-mediated transitions are balanced by progesterone (P4). We will test our hypotheses using 2 specific aims: Aim 1—To determine if LPS (infection) and TNFα (inflammation) can cause TGFβ -TAB1-p38MAPK signaling pathways, leading to EMT in AEC. Aim 2—To determine the regulatory role of P4 in LPS and TNFα-mediated TGFβ production and its ability to reduce EMT. Maintenance of membrane homeostasis by TGFβ-P4 during normal conditions and its disruption by infection/inflammation promoting EMT may provide a novel pathway to preterm parturition mediated by membrane disruption.

抽象的 胎膜（羊膜）提供结构框架并执行机械和保护作用 怀孕期间的功能。渐进性 p38 丝裂原激活蛋白激酶 (MAPK) 介导的 衰老（衰老机制）发生在胎膜中。它与胎儿的生长发育有关， 由于宫腔内氧化应激（OS）增加，足月时加速。胎儿衰老 组织引起炎症，促进分娩；因此，膜在怀孕和怀孕过程中起着至关重要的作用 分娩。在整个妊娠过程中，细胞膜通过自我修复来维持体内平衡。 脱落并且基质降解。这种重塑过程会产生间隙和微裂缝。在蜂窝 水平，目前尚不清楚膜的多能羊膜上皮细胞和间充质细胞如何修复 受损地点。由于重塑对于维持膜稳态和防止不良反应至关重要 妊娠结果，了解这个过程至关重要。我们确定羊膜损伤 上皮细胞（AEC）迫使AEC增殖和细胞迁移以重建受损部位，这是正常的 生理反应。试验数据表明 OS 和 p38 MAPK 介导的衰老导致上皮细胞 间充质转化 (EMT) 并通过增加 TGFβ 的产生来防止组织重塑。我们也 确定 TGFβ 具有双重功能：1）通过 p38MAPK 自身磷酸化诱导衰老 2) 诱导 EMT 转录因子以促进细胞膜和机械膜的破坏。这些 黄体酮（P4）可以逆转这种影响。我们假设平衡的组织重塑维持胎儿 妊娠期间膜稳态，但由于足月或反应压倒性 OS 导致 TGFβ 增加 早产时感染和炎症 (TNFα) 导致 p38MAPK 介导的不可逆状态 衰老和 EMT 会促进膜损伤。此外，我们假设在 怀孕期间，TGFβ 介导的转变由黄体酮 (P4) 平衡。我们将使用以下方法来检验我们的假设 2具体目标： 目标 1 — 确定 LPS（感染）和 TNFα（炎症）是否会导致 TGFβ -TAB1-p38MAPK 信号通路，导致 AEC 中的 EMT。 目标 2——确定 P4 在 LPS 和 TNFα 介导的 TGFβ 产生中的调节作用及其能力 以减少 EMT。正常情况下TGFβ-P4对膜稳态的维持及其作用 感染/炎症破坏促进 EMT 可能为早产提供新途径 由膜破坏介导。