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，羊水（AF）容量异常发生在5-10%的妊娠中，并且通常与母体和胎儿发病率增加相关，从而损害妊娠结局并产生潜在的长期不良后果。这大大增加了对临床干预的需求，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，尽管发病率很高，仍然没有有效的治疗方法来纠正不适当的房颤容量，从而提高新生儿存活率。这主要是由于缺乏对调节心房颤动量的机制和最终导致心房颤动量过大或减少的异常原因的理解。关于这一主题的研究数量有限，主要是因为在没有适当的人体模型的情况下，很难设计相对未知和复杂的调节机制的研究。实验动物的研究已经产生了目前关于AF在羊膜（运输的限速层）中的运输特性的大部分知识。然而，一个关键的问题仍然是这些动物数据对人类的适用性，以及使用动物模型进行人类生物学研究的相关性。为了解决这一不足，我们建议建立一个研究心房颤动体积调节的人体模型。我们最近开发了一种体外羊羊膜细胞模型，用于研究AF在羊膜细胞单层上的运输，该模型易于实验操作。我们的目标是适应这种细胞培养系统运输研究在人羊膜细胞。本应用程序旨在研究该体外人羊膜细胞模型在正常妊娠、羊水过少和羊水过多患者AF转运机制研究中的有效性。在Specific Aim 1中，将确定AF在人羊膜细胞单层中的主动和被动转运成分作为溶质分子量的函数；小泡在主动跨细胞运输中的作用将被研究；探讨VEGF165及其抑制异构体在调节主动运输中的作用；并将阐明羊水过少和羊水过多时羊膜细胞运输特性的改变。在特异性目标2中，将研究VEGF165对人羊膜细胞中水通道蛋白1、3和9的被动扩散的调节。我们将分析羊水过少和羊水过多是否会改变羊水通道蛋白在羊膜细胞中的被动扩散。在Specific Aim 3中，我们将在正常妊娠、羊水过少和羊水过多的羊膜细胞中研究VEGF165作为细胞信号传导事件的调节因子在介导活跃性小泡胞吞作用中的作用。综上所述，本研究将揭示该人羊膜细胞培养模型是否适合用于研究人类妊娠期心房纤颤体积调节机制，并验证该模型用于解读羊水过少和羊水过多患者心房纤颤体积异常病因的可行性。