Human Amnion Cell Model for Investigation of Amniotic Fluid Volume

用于研究羊水量的人类羊膜细胞模型

• 批准号: 8113659
• 负责人: CECILIA CHEUNG
• 金额: $ 7.7万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8113659
• 关键词: 1-Phosphatidylinositol 3-Kinase; AQP1 gene; Active Biological Transport; Address; Amniotic Fluid; Animal Model; Animals; Caveolae; Cell Culture System; Cell Culture Techniques; Cell model; Cells; Characteristics; Clinical; Complex; Data; Diffusion; Environment; Etiology; Event; Fetal Growth; Goals; Human; Human Biology; In Vitro; Incidence; Intervention; Investigation; Knowledge; Lead; Liquid substance; Maintenance; Mediating; Mediation; Membrane Microdomains; Modeling; Molecular Weight; Morbidity - disease rate; Neonatal; Normal Cell; Oligohydramnios; Pathway interactions; Perinatal; Personal Satisfaction; Polyhydramnios; Pregnancy; Pregnancy Outcome; Pregnancy Tests; Process; Property; Protein Isoforms; Protein Kinase; Protein Kinase C; Regulation; Research; Research Design; Role; SRC gene; Sheep; Signal Pathway; Signal Transduction; Signal Transduction Pathway; Sister; Structural Protein; Study models; System; Testing; Therapeutic; Transport Process; Treatment Protocols; Urea; VEGF165; Validation; Vascular Endothelial Growth Factor Receptor; Vascular Endothelial Growth Factor Receptor-2; Vascular Endothelial Growth Factors; adverse outcome; amnion; animal data; aquaporin 3; aquaporin 4; base; caveolin 1; cost; design; effective therapy; fetal; improved; interest; monolayer; passive transport; pregnant; receptor; solute; transcytosis; water channel

DESCRIPTION (provided by applicant): Abnormalities in amniotic fluid (AF) volume occur in 5-10% of pregnancies in the US and are often associated with increased maternal and fetal morbidity that compromises pregnancy outcome with potential long term adverse consequences. This dramatically increases the need for clinical intervention and thus costs of obstetrical management. Approximately 50% of symptomatic oligohydramnios and polyhydramnios have unknown etiology. However despite the high incidence, there remains no effective therapy for correcting the inappropriate AF volume thereby improving neonatal survival. This is primarily due to the lack of understanding of the mechanisms that regulate AF volume and the causes of the abnormalities that ultimately lead to excessive or reduced AF volumes. The limited amount of research on this subject is largely because of the difficulties in designing studies on relatively unknown and complex regulatory mechanisms without an appropriate human model. Studies in experimental animals have generated most of the current knowledge on AF transport characteristics across the amnion, the rate-limiting layer for transport. However, a critical question remains as to the applicability of these animal data to the human, and the relevance of using animal models for investigations of human biology. To address this deficiency, we propose to develop a human model for studies of AF volume regulation. We have recently developed an in vitro ovine amnion cell model for the study of AF transport across amnion cell monolayers that are readily accessible to experimental manipulations. Our goal is to adapt this cell culture system for transport studies in human amnion cells. The present application is designed to investigate the validity of this in vitro human amnion cell model for studies of AF transport mechanisms in normal pregnancies and in oligohydramnios and polyhydramnios. In Specific Aim 1, the active and passive components of AF transport across human amnion cell monolayers as a function of solute molecular weight will be determined; the involvement of caveolae in active transcellular transport will be investigated; the role of VEGF165 and its inhibitory isoform in regulating active transport will be explored; and alterations in transport characteristics in amnion cells from oligohydramnios and polyhydramnios will be elucidated. In Specific Aim 2, the VEGF165 regulation of passive diffusion across water channels aquaporins 1, 3 and 9 in human amnion cells will be examined. Whether passive diffusion across aquaporin channels in amnion cells from oligohydramnios and polyhydramnios is altered will be analyzed. In Specific Aim 3, the role of VEGF165 as the regulatory factor in the cell signaling events that mediate active caveolar transcytosis will be investigated in amnion cells from normal pregnancy, and from oligohydramnios and polyhydramnios. Overall, the studies will reveal whether this human amnion cell culture model is a suitable model for the investigation of AF volume regulatory mechanisms in human pregnancy, and test the feasibility of using this model to decipher the etiology underlying abnormal AF volume in oligohydramnios and polyhydramnios. PUBLIC HEALTH RELEVANCE: The proposed studies will establish and characterize a human amnion cell model for the investigation of mechanisms mediating amniotic fluid volume regulation in human pregnancy. Knowledge gained from studies using this model will allow the elucidation of the etiology of oligohydramnios and polyhydramnios, and thus provide the scientific basis for designing management protocols and treatment paradigms for amniotic fluid abnormalities.

描述（由申请人提供）：在美国，5-10% 的妊娠发生羊水 (AF) 量异常，并且通常与母体和胎儿发病率增加相关，从而影响妊娠结局，并可能产生长期不良后果。这极大地增加了临床干预的需求，从而增加了产科管理的成本。大约 50% 的症状性羊水过少和羊水过多病因不明。然而，尽管发生率很高，但仍然没有有效的治疗方法来纠正不适当的房颤体积，从而提高新生儿的生存率。这主要是由于缺乏对调节 AF 容量的机制以及最终导致 AF 容量过多或减少的异常原因的了解。关于这一主题的研究数量有限，很大程度上是因为在没有适当的人体模型的情况下，设计相对未知和复杂的调节机制的研究很困难。对实验动物的研究已经产生了关于 AF 跨羊膜（传输的限速层）传输特性的大部分最新知识。然而，一个关键问题仍然是这些动物数据对人类的适用性，以及使用动物模型研究人类生物学的相关性。为了解决这一缺陷，我们建议开发一个用于研究房颤音量调节的人体模型。我们最近开发了一种体外绵羊羊膜细胞模型，用于研究 AF 跨羊膜细胞单层的转运，该模型易于实验操作。我们的目标是使该细胞培养系统适用于人类羊膜细胞的运输研究。本申请旨在研究该体外人类羊膜细胞模型对于正常妊娠以及羊水过少和羊水过多中 AF 转运机制研究的有效性。在具体目标 1 中，将确定 AF 跨人类羊膜细胞单层转运的主动和被动成分，作为溶质分子量的函数；将研究小窝参与主动跨细胞运输；将探讨 VEGF165 及其抑制亚型在调节主动转运中的作用；将阐明羊水过少和羊水过多引起的羊膜细胞运输特性的变化。在具体目标 2 中，将检查 VEGF165 对人羊膜细胞中水通道蛋白 1、3 和 9 被动扩散的调节。将分析羊水过少和羊水过多时羊膜细胞中水通道蛋白通道的被动扩散是否发生改变。在具体目标 3 中，将在正常妊娠、羊水过少和羊水过多的羊膜细胞中研究 VEGF165 作为调节因子在介导活性小窝转胞吞作用的细胞信号传导事件中的作用。总体而言，这些研究将揭示这种人类羊膜细胞培养模型是否适合研究人类妊娠期房颤体积调节机制，并测试使用该模型破译羊水过少和羊水过多时房颤体积异常病因的可行性。 公共健康相关性：拟议的研究将建立并表征人类羊膜细胞模型，用于研究人类妊娠期间羊水量调节的调节机制。从使用该模型的研究中获得的知识将有助于阐明羊水过少和羊水过多的病因，从而为设计羊水异常的管理方案和治疗范例提供科学依据。