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

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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.