Novel 3-D Model of Placental Cytomegalovirus Infection

胎盘巨细胞病毒感染的新型 3D 模型

• 批准号: 6718306
• 负责人: CINDY Anne MORRIS
• 金额: $ 7.43万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-12-01 至 2005-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6718306
• 关键词: Herpesviridae disease; bioreactors; biotechnology; cell aggregation; cell differentiation; cell line; cell proliferation; clinical research; cytomegalovirus; fluorescence microscopy; gene expression; host organism interaction; immunofluorescence technique; keratin; model design /development; monoclonal antibody; placenta; pregnancy infection; trophoblast; virus cytopathogenic effect; women' s health

DESCRIPTION (provided by applicant): The overall objective of this proposal is to develop an improved tissue model for the study of human cytomegalovirus infection (CMV) during pregnancy. Specifically, rotating wall vessels (RWVs) will be utilized to establish this advanced model system. CMV is the leading cause of congenital viral infection, and intrauterine transmission of the virus occurs in approximately 50% of pregnant women with primary CMV infection. CMV infection during pregnancy can have catastrophic consequences not only on the neonate, but also on pregnancy outcome, including spontaneous abortion, intrauterine growth restriction and pre-eclampsia. Despite the significant morbidity and mortality associated with congenital CMV infection, little is known about how the virus infects the conceptus. RWV technology offers a unique approach not previously applied toward the study of placental tissues and provides a novel model system to explore intrauterine CMV infection. The culture conditions of these bioreactors provide a low-shear and low-turbulence growth environment, similar to that required for normal in vivo fetal and placental development. Such an environment allows cells to co-localize spatially, grow three-dimensionally, and differentiate into highly specialized tissues. Preliminary data presented in this proposal demonstrate that the RWV supports the growth of a first trimester cytotrophoblast cell line. Importantly, we have shown that the environment created within the bioreactor promotes cytotrophoblast differentiation and allows for the formation of complex, multi-layered three-dimensional (3-D) aggregates that are functionally similar to in utero trophoblast cells. The hypothesis to be tested within this proposal is that placental epithelial cells cultured in the RWV are functionally similar to the developing in utero placenta and that the 3-D growth of these cells will provide an improved model system for the study of the molecular details involved in intrauterine CMV infection. The long-term research goals of this proposal are to utilize the 3-D model to study and understand 1) the molecular details of normal placental development, 2) CMV pathogenesis, 3) host-pathogen interactions that occur at the maternal-fetal interface during pregnancy, and 4) the molecular mechanism(s) that regulate the effect of CMV on placentation and pregnancy outcome. The specific aims are: 1) to further characterize the 3-D model of human placentation, and 2) to apply the placental 3-D model toward the study of intrauterine CMV infection. This novel model system will contribute to advanced studies of human placentation, ultimately leading to new therapeutic strategies for the prevention and treatment of placental and congenital CMV infection.

描述（由申请人提供）：该提案的总体目的是开发改进的组织模型，以研究怀孕期间人类巨细胞病毒感染（CMV）。具体而言，将利用旋转的壁船（RWV）来建立此高级模型系统。 CMV是先天性病毒感染的主要原因，病毒的宫内传播发生在大约50％的原发CMV感染的孕妇中。怀孕期间的CMV感染不仅会对新生儿产生灾难性后果，而且还会对怀孕结局产生灾难性的后果，包括自发流产，宫内内生长限制和先兆子痫。尽管与先天性CMV感染相关的发病率和死亡率很大，但对病毒如何感染该概念知之甚少。 RWV技术提供了一种以前不适用于研究胎盘组织的独特方法，并提供了一种新型的模型系统来探索宫内CMV感染。这些生物反应器的培养条件提供了低剪切和低扰动的生长环境，类似于正常体内胎儿和胎盘发育所需的环境。这样的环境使细胞可以在空间上共定位，在三维生长并分化为高度专业的组织。该提案中介绍的初步数据表明，RWV支持头三个月的细胞增生细胞细胞系的生长。重要的是，我们已经表明，生物反应器中创建的环境促进了细胞增生细胞分化，并允许形成与子宫滋养细胞在功能上相似的复杂的，多层的三维（3-D）聚集体。在该提案中要检验的假设是，在RWV中培养的胎盘上皮细胞在功能上与子宫胎盘中的发育相似，并且这些细胞的3-D生长将为研究涉及诊所内CMV感染的分子细节的研究提供改进的模型系统。该提案的长期研究目标是利用3-D模型来研究和理解1）正常胎盘发育的分子细节，2）CMV发病机理，3）3）宿主 - 病原体相互作用，在孕妇界面上发生的宿主 - 病原体相互作用，以及4）4）分子机制（S）调节CMV对置换和怀孕的影响。具体目的是：1）进一步表征人体胎盘的3-D模型，以及2）将胎盘3-D模型应用于研究宫内CMV感染的研究。这种新型模型系统将有助于对人体胎盘的高级研究，最终导致新的治疗策略，以预防和治疗胎盘和先天性CMV感染。