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

绒毛滋养细胞增殖、分化和凋亡的平衡是维持胎母界面功能和形态完整的保证。关于它的调节，所有这些过程都是紧密相连的，它们由各种细胞因子调节。这种失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重要意义。绒毛滋养细胞强烈表达的生长激素前颗粒蛋白，在子痫前期和胎儿生长受限的情况下胎盘表达增加。本研究旨在探讨前颗粒蛋白对绒毛滋养细胞再生、生长、分化和凋亡的功能影响。此外，我们将探讨滋养细胞前颗粒蛋白调控的两种潜在相关机制。首先，我们打算评估miR-195对前颗粒蛋白的转录后调控，miR-195对前颗粒蛋白mRNA具有特异性亲和力，对许多参与细胞凋亡和增殖的蛋白质的调控具有重要意义。其次，我们的研究将重点关注缺氧，即氧化应激对颗粒前蛋白表达的影响。鉴于这些因素与子痫前期及胎儿生长受限的相关性，应进一步探讨前颗粒蛋白调控的滋养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，如原代分离的滋养细胞、胎盘外植体、bewo细胞或HUVECs。滋养层细胞的病毒转导会导致靶基因过表达或敲低，从而在实验中修饰前颗粒蛋白和miR-195的表达。将通过功能测试以及转录组和蛋白表达分析进一步分析连续效应。收集到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。

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.