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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.

绒毛滋养层细胞增殖、分化和凋亡的平衡保证了母胎界面功能和形态的完整性。就其调节而言，所有这些过程都是密切相关的，并且它们受到各种细胞因子的调节。不平衡对于妊娠相关疾病如先兆子痫或胎儿生长受限的发展具有突出的意义。生长激素颗粒蛋白前体，这是强烈表达的绒毛滋养层细胞，揭示了增加胎盘表达的情况下，先兆子痫以及胎儿生长受限。本研究的目的是探讨颗粒蛋白前体对绒毛滋养层细胞再生、生长、分化和凋亡的功能影响。此外，我们将研究两个潜在的相关机制的滋养层颗粒蛋白前体的调节。首先，我们打算评估miR-195对颗粒蛋白前体的转录后调节，miR-195对颗粒蛋白前体mRNA具有特异性亲和力，并且对参与凋亡和增殖的许多蛋白质的调节具有重要意义。其次，我们的研究将集中在缺氧，分别氧化应激，颗粒蛋白前体表达的影响。关于这些因素与先兆子痫和胎儿生长受限的相关性，应探讨颗粒蛋白前体调节的滋养层细胞对内皮细胞的影响。对于实验设置，将使用适当的细胞培养模型，如原代分离的滋养层细胞、胎盘外植体、BeWo细胞或HUVEC。颗粒蛋白前体和miR-195的表达将通过滋养层细胞的病毒转导进行实验修饰，导致靶基因的过表达或敲低。将通过功能测试以及转录组和蛋白质表达的分析来进一步分析连续效应。收集的见解可以更好地理解妊娠相关疾病的发病机制，并可能提供治疗方法。