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Placental Endogenous Retrovirus Expression

胎盘内源性逆转录病毒表达

基本信息

批准号：
6694047
负责人：
NEAL STEWART ROTE
金额：
$ 7.58万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-02-01 至 2006-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6694047
关键词：
Retroviridae biomaterial development /preparation cell differentiation cell line clinical research enzyme linked immunosorbent assay human tissue immunocytochemistry laboratory mouse messenger RNA monoclonal antibody placenta polymerase chain reaction technology /technique development tissue /cell culture transfection trophoblast virus protein

项目摘要

DESCRIPTION (provided by applicant): Approximately 1% of the human genome appears to be of retroviral origin. The purpose of these "endogenous" retroviral elements in human development remains unknown. A small number of organs express endogenous retroviruses under physiologic conditions; the human placenta is one of these. The normal human placental villous cytotrophoblast expresses differentiation-related transcription of the env region of a single copy endogenous retroviral genome (ERV-3). The strong association of ERV-3 expression with normal trophoblast differentiation suggests a physiologic role for ERV-3 in placental development. In support of this hypothesis, are the following observations. 1) ERV-3 env mRNA and Env protein are increased in syncytiotrophoblast and in spontaneously differentiating mononuclear villous cytotrophoblast. 2) Differentiation-related increase in ERV-3 env mRNA is concurrent with increased b-hCG mRNA and protein and intercellular fusion. 3) Transfection of BeWo choriocarcinoma cells, a model for trophoblast differentiation, with the sense strand of ERV-3 env resulted in a stable transfectant in which most aspects of trophoblast differentiation occurred: decreased growth, morphologic differentiation, and increased b-hCG mRNA and hCG protein production. 4) Transfection with the antisense strand decreased b-hCG mRNA production when the cells were induced to differentiate. These preliminary data support a hypothesis that ERV-3 env expression is an important step in the pathway of normal trophoblast differentiation, potentially through regulation of hCG expression. The specific aim of this R03 application is to accumulate adequate preliminary data for a successful R01 application to test this hypothesis, The individual specific Aims are: Aim 1) is to develop reagents, e.g.,monoclonal antibody and cell lines transfected with inducible plasmid systems, for studying the role of ERV3 env in villous trophoblast differentiation; Aim 2) is to determine whether ERV-3 env expression is necessary for differentiation of freshly isolated villous cytotrophoblasts; and Aim 3) is to determine whether ERV-3 env expression is inducible by steroids. These specific aims were chosen to address issues raised in previously unsuccessful R01 applications.

描述（由申请人提供）：大约1%的人类基因组似乎是逆转录病毒来源的。这些“内源性”逆转录病毒成分在人类发育中的目的仍然未知。少数器官在生理条件下表达内源性逆转录病毒;人胎盘是其中之一。正常人胎盘绒毛细胞滋养层表达单拷贝内源性逆转录病毒基因组（ERV-3）的env区的分化相关转录。ERV-3表达与正常滋养层分化的强相关性表明ERV-3在胎盘发育中的生理作用。为了支持这一假设，有以下观察结果。1)ERV-3 env mRNA和Env蛋白在合体滋养细胞和自发分化的单核绒毛细胞滋养细胞中增加。2)ERV-3 env mRNA的分化相关增加与b-hCG mRNA和蛋白以及细胞间融合的增加同时发生。3)转染BeWo绒毛膜癌细胞，滋养层分化的模型，与ERV-3 env的正义链导致在一个稳定的转染子中，滋养层分化的大部分方面发生：生长下降，形态分化，并增加b-hCG mRNA和hCG蛋白的生产。4)当细胞被诱导分化时，用反义链转染降低b-hCG mRNA的产生。这些初步的数据支持一个假设，ERV-3 env表达是正常滋养层细胞分化途径中的一个重要步骤，可能通过调节hCG表达。本R 03申请的具体目的是为成功的R 01申请积累足够的初步数据，以检验该假设。各个具体目的是：目的1）是开发试剂，例如，单克隆抗体和用诱导型质粒系统转染的细胞系，用于研究ERV-3 env在绒毛滋养层细胞分化中的作用;目的2）是确定ERV-3 env表达是否是新鲜分离的绒毛细胞滋养层细胞分化所必需的;和目的3）是确定ERV-3 env表达是否可由类固醇诱导。选择这些具体目标是为了解决以前不成功的R 01申请中提出的问题。