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: 8242678
• 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 2014-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8242678
• 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; public health relevance; 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)量异常发生在5%-10%的怀孕中，通常与孕妇和胎儿发病率的增加有关，这会危及妊娠结果，并可能产生长期的不良后果。这极大地增加了临床干预的需要，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的病因不明。然而，尽管发病率很高，但仍然没有有效的治疗方法来纠正不适当的房颤体积，从而提高新生儿存活率。这主要是由于对房颤量的调节机制和最终导致房颤量过多或减少的异常的原因缺乏了解。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，很难设计关于相对未知和复杂的监管机制的研究。对实验动物的研究已经产生了目前关于房颤通过羊膜的运输特性的大部分知识，羊膜是运输的限速层。然而，关键的问题仍然是这些动物数据对人类的适用性，以及使用动物模型研究人类生物学的相关性。为了解决这一不足，我们建议建立一个人体模型来研究房颤的容量调节。我们最近建立了一个体外羊膜细胞模型，用于研究房颤通过羊膜细胞单层的转运，该模型易于进行实验操作。我们的目标是使这种细胞培养系统适用于人羊膜细胞的运输研究。本研究旨在探讨这一体外人羊膜细胞模型在研究正常妊娠、羊水过少和羊水过多时房颤转运机制的有效性。在具体目标1中，将确定房颤跨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；将研究小凹在主动跨细胞转运中的参与；将探索VEGF165及其抑制亚型在调节主动转运中的作用；并将阐明羊水过少和羊水过多时羊膜细胞转运特性的变化。在具体目标2中，将研究VEGF165对人羊膜细胞水通道水通道蛋白1、3和9被动扩散的调节。羊水过少和羊水过多羊膜细胞水通道被动扩散是否发生改变，有待进一步研究。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作为调节因子在细胞信号事件中的作用，这些信号事件介导了活动性空泡穿越细胞。总之，这些研究将揭示这种羊膜细胞培养模型是否适合于研究人类妊娠时房颤的容量调节机制，并测试使用该模型来解释羊水过少和羊水过多的房颤容量异常的病因的可行性。 公共卫生相关性：拟议的研究将建立和表征一个人羊膜细胞模型，用于研究调节人类妊娠期间羊水量调节的机制。利用该模型所获得的知识将有助于阐明羊水过少和羊水过多的病因，从而为制定羊水异常的处理方案和治疗方案提供科学依据。