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）：美国5-10％的怀孕期间发生羊水（AF）的异常，并且通常与孕产妇和胎儿发病率增加有关，从而损害了怀孕，并带来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％的症状性寡素体和多氢化植物具有未知的病因。然而，尽管发病率很高，但仍未有效纠正不适当的AF量，从而改善新生儿生存率。这主要是由于缺乏对调节AF量的机制的了解以及最终导致过度或减少AF量的异常原因的原因。对这一主题的研究有限，主要是因为在没有适当的人类模型的情况下设计了对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整个羊膜中AF运输特征的大多数知识，该羊膜是运输速率的限制层。但是，关于这些动物数据对人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仍然存在一个关键问题。为了解决这一缺陷，我们建议开发一个人类模型，以研究AF量调节。我们最近开发了一种体外卵巢羊膜细胞模型，用于研究跨羊膜细胞单层的AF转运，这些单层很容易被实验操作访问。我们的目标是使该细胞培养系统适应人类羊膜细胞中的运输研究。本应用旨在研究该体外人羊膜细胞模型的有效性，用于研究正常妊娠，少聚物化热和多氢化物中的AF转运机制。在特定的目标1中，将确定AF转运的主动和被动成分横跨人羊膜细胞单层作为溶质分子量的函数。将研究口腔参与活性的跨细胞运输； VEGF165及其抑制同工型在调节主动运输中的作用；将阐明来自寡氢化和多氢化物的羊膜细胞中转运特性的改变。在特定的目标2中，将检查VEGF165在人羊膜细胞中对水通道1、3和9跨水通道的被动扩散的调节。是否会分析是否会改变少聚物杂种和多氢化物的羊膜通道的被动扩散。在特定的目标3中，VEGF165作为调节因子在细胞信号事件中的作用，将在正常妊娠的羊膜细胞中，以及少血离杂志和多化合物中调查介导活性洞穴跨性化的调节因子。总体而言，研究将揭示这种人类羊膜细胞培养模型是否是研究人类怀孕中AF体积调节机制的合适模型，并测试使用该模型破译寡素体和多hyhydramnios中基本异常AF量的病因的可行性。 公共卫生相关性：拟议的研究将建立并表征人类羊膜细胞模型，以研究介导人妊娠中赋予羊水体积调节的机制。使用该模型从研究中获得的知识将允许阐明寡氢化植物和多氢化物的病因，因此为设计管理方案和治疗范式提供了科学基础，以赋予羊水异常。