Analysis of function and regulation of progranulin and miR-195 on human villous trophoblast cells and its impact on endothelial function

颗粒体蛋白前体和miR-195对人绒毛滋养层细胞的功能调控及其对内皮功能的影响分析

基本信息

批准号：
250054264
负责人：
Professorin Dr. Brigitte M. Pützer
金额：
--
依托单位：
Universitätsfrauenklinik und Poliklinik am Klinikum Südstadt Rostock
依托单位国家：
德国
项目类别：
Research Grants
财政年份：
2014
资助国家：
德国
起止时间：
2013-12-31 至 2017-12-31
项目状态：
已结题

来源：
https://gepris.dfg.de/gepris/projekt/250054264?language=en
关键词：
Analysis function regulation progranulin miR

项目摘要

The balance of proliferation, differentiation and apoptosis of villous trophoblast cells guarantees the maintenance of the functional and morphological integrity of the fetal-maternal interface. With regard to its regulation all of these processes are closely connected and they are regulated by miscellaneous cytokines. An imbalance is of outstanding significance for the development of pregnancy related diseases like preeclampsia or fetal growth restriction. The growth hormone progranulin, which is strongly expressed by villous trophoblast cells, revealed an increased placental expression in cases of preeclampsia as well as fetal growth restriction. The aim of this study will be the exploration of the functional impact of progranulin on regeneration and growth as well as differentiation and apoptosis of villous trophoblast cells. Furthermore, we will investigate two potentially relevant mechanisms of the trophoblastic progranulin regulation. Firstly, we intend to evaluate the posttranscriptional regulation of progranulin by miR-195, which exhibits a specific affinity to progranulin mRNA and is significant for the regulation of numerous proteins involved in apoptosis and proliferation. Secondly, our study will focus on the impact of hypoxia, respectively oxidative stress, on progranulin expression. With respect to the relevance of these factors for the development of preeclampsia and fetal growth restriction, the effect of progranulin regulated trophoblast cells on endothelial cells should be explored. For the experimental setup appropriate cell culture models like primary isolated trophoblast cells, placental explants, BeWo-cells or HUVECs will be used. The expression of progranulin and miR-195 will be experimentally modified by viral transduction of trophoblast cells resulting in an overexpression or knock down of the target gene. The consecutive effects will be further analyzed by functional tests as well as analysis of the transcriptome and protein expression. Gathered insights could account for a better understanding of pathogenesis of pregnancy related diseases and may offer therapeutic approaches.

绒毛细胞细胞的增殖，分化和凋亡的平衡可确保维持胎儿婚礼的功能和形态完整性。关于其调节，所有这些过程都密切相关，并且由其他细胞因子调节。不平衡对于妊娠相关疾病（如先兆子痫或胎儿生长限制）具有重要意义。由绒毛细胞细胞强烈表达的生长激素前素蛋白在先兆子痫和胎儿生长限制的情况下显示出胎盘表达的增加。这项研究的目的是探索尿素对再生和生长的功能影响以及绒毛细胞细胞的分化和凋亡。此外，我们将研究滋养细胞促进蛋白调节的两种潜在相关机制。首先，我们打算评估MiR-195对尿素蛋白的转录后调节，该调节表现出对progranulin mRNA的特定亲和力，对于调节众多参与凋亡和增殖的众多蛋白质非常重要。其次，我们的研究将重点介绍缺氧分别对氧化应激的影响，对前粒蛋白表达。关于这些因素与先兆子痫和胎儿生长限制的相关性，应探讨progranulin调节的滋养细胞对内皮细胞的影响。对于实验性设置，将使用适当的细胞培养模型，例如原发性分离的滋养细胞细胞，胎盘外植体，bewo细胞或HUVEC。促素蛋白和miR-195的表达将通过病毒转导的滋养细胞细胞进行实验修饰，从而导致过表达或敲低靶基因。连续效应将通过功能测试以及转录组和蛋白质表达的分析进一步分析。收集的见解可以更好地理解与妊娠相关疾病的发病机理，并可能提供治疗方法。