Intercellular interactions define cell migrations and transitions that maintain fetal membrane homeostasis

细胞间相互作用定义了维持胎膜稳态的细胞迁移和转变

• 批准号: 10356919
• 负责人: Arum Han
• 金额: $ 43.99万
• 依托单位: UNIVERSITY OF TEXAS MED BR GALVESTON
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-12 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10356919
• 关键词: Aging; Amniotic Fluid; Area; Basement membrane; Biological; Biological Assay; Biological Models; Biomedical Engineering; Birth; Cell Communication; Cell Culture Techniques; Cell Shape; Cell membrane; Cell model; Cell physiology; Cells; Characteristics; Development; Devices; Environment; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Exhibits; Experimental Models; Exposure to; Extracellular Matrix; Failure; Fetal Membranes; Fetus; Fissural; Functional disorder; Generations; Homeostasis; Impairment; In Situ; In Vitro; Infection; Inflammation; Inflammatory; Intervention; Investigation; Kinetics; Knowledge; Label; Laboratories; Lead; Magnetism; Mechanics; Membrane; Mesenchymal; Methodology; Methods; Microfluidics; Microscopy; Modeling; Molecular; Monitor; Oxidative Stress; Pathologic; Pathway interactions; Physiological Processes; Pregnancy; Pregnancy Complications; Premature Birth; Process; Property; Proteins; Recycling; Research; Risk; Role; Signal Transduction; Site; System; Technology; Testing; Time; Tissues; Transitional Epithelium; Translating; Urea; Uterine cavity; Width; amnion; bone fracture repair; cell motility; cell type; design; epithelial to mesenchymal transition; fetus cell; healing; improved; in utero; in vitro Model; innovation; membrane model; migration; morphometry; multidisciplinary; organ on a chip; premature; preterm premature rupture of membranes; prototype; repaired; response; risk minimization; seal; second harmonic; senescence

Fetal membranes (amniochorion) protect the fetus during pregnancy. At term, senescence (aging) and inflammation cause functional and mechanical instability to membrane cells, contributing to parturition. Premature senescence and membrane dysfunctions are associated with preterm birth (PTB) and preterm premature rupture of the membranes (pPROM). However, cellular-level changes contributing to membrane stability during gestation and its dysfunction leading to labor and delivery are still unclear. Recent studies of senescent term and preterm membranes revealed “microfractures” (MFs), sites of cellular remodeling. MFs are resealed during gestation to maintain membrane integrity. Higher numbers of MFs and their increased morphometry (depth and width) in term labor, pPROM, and PTB membranes compared to respective controls suggest MFs' resealing is compromised. Amnion epithelial cells in MFs have been observed undergoing epithelial mesenchymal transition (EMT). Further, these cells showed proliferative and resealing properties of deepithelialized (nude/cell free) areas to stabilize membranes. At the healing edge of MFs, amnion mesenchymal cells exhibited a reverse phenomenon, mesenchymal-epithelial transition (MET). From these findings, we postulate that cellular transitions are essential for maintaining fetal membrane integrity. We hypothesize that MFs are areas of membrane remodeling and their increased number and morphometry are associated with failure to remodel and dysfunctional membranes. Understanding intercellular and cell-matrix interactions causing MFs' development and their resealing will help us to determine how oxidative stress (OS) and inflammation can contribute to the persistence of MFs and dysfunctional membrane status in PTB and pPROM. Two specific aims to be tested are Specific Aim 1: To investigate the dynamic remodeling of the fetal membrane epithelium in an in vitro model of cell-free (nude) membranes during OS and infection / inflammation compared to normal conditions; Specific Aim 2: To determine cell migration, matrix degradation, and cellular transition associated with MFs' formation using a fetal membrane organ-on-a-chip approach. This multidisciplinary proposal combines cell and molecular biological and bioengineering approaches designed to overcome the limitations of classic 2D cell cultures by developing a fetal membrane-on-a-chip using organ-on-chip technologies. This model system will maintain multiple cell types in close proximity with constant dynamic interactions, similar to the conditions in utero. We will elucidate causative molecular mechanisms of (normal and abnormal) biologic MFs' formation and how they contribute to PTB and pPROM. Understanding cellular-level mechanisms will allow us to design strategies to minimize MFs' development to strengthen intrauterine cavities and reduce the risk of PTB and pPROM.

胎膜（绒毛膜）在怀孕期间保护胎儿。在足月时，衰老（老化）和炎症导致膜细胞功能和机械不稳定，有助于分娩。早衰和膜功能障碍与早产（PTB）和早产胎膜早破（pPROM）有关。然而，细胞水平的变化，有助于膜稳定性在妊娠期间，其功能障碍，导致劳动和交付仍然不清楚。最近对衰老足月和早产儿胎膜的研究揭示了“微裂缝”（MF），即细胞重塑的部位。在妊娠期间重新密封MF以保持膜的完整性。与相应对照相比，足月分娩、pPROM和PTB膜中MF数量更高及其形态测定学（深度和宽度）增加表明MF的重新密封受到损害。已观察到MF中的羊膜上皮细胞经历上皮间质转化（EMT）。此外，这些细胞显示出增殖和深上皮化（裸/无细胞）区域的重新密封特性，以稳定膜。在MF愈合边缘，羊膜间充质细胞表现出相反的现象，间充质上皮转化（MET）。从这些发现，我们假设，细胞的转变是必不可少的维持胎膜的完整性。我们 假设MF是膜重塑的区域，并且它们的数量和形态学增加与膜重塑失败和功能障碍有关。理解细胞间和细胞基质 引起微纤维发展和重新密封的相互作用将有助于我们确定氧化应激（OS） 并且炎症可导致MF的持续存在和PTB和pPROM中的膜功能障碍状态。待测试的两个具体目的是具体目的1：研究与正常条件相比，在OS和感染/炎症期间无细胞（裸）膜体外模型中胎膜上皮的动态重塑;具体目的2：使用胎膜器官芯片方法确定与MF形成相关的细胞迁移、基质降解和细胞转化。这个多学科的建议结合了细胞和分子生物学和生物工程的方法，旨在克服传统的二维细胞培养的局限性，通过开发一个胚胎膜芯片上使用器官芯片技术。该模型系统将保持多种细胞类型紧密接近，具有恒定的动态相互作用，类似于子宫内的条件。我们将阐明（正常和异常）生物MFs形成的分子机制，以及它们如何导致PTB和pPROM。了解细胞水平的机制将使我们能够设计策略，以最大限度地减少MF的发展，以加强宫腔和降低PTB和pPROM的风险。