Embryonal non-coding RNAs in the human placenta and the maternal circulation.

非编码RNA(NcRNAs)在几乎所有的细胞、器官和物种中都发挥着中心和调节的作用，这一点自人类ENCODE计划和其他几个基因组计划以来得到了广泛的认识。然而，我们只知道一小部分ncRNA。此外，长的非编码RNA只有大约五年的历史，在胎盘中的研究非常少。到目前为止，大多数已被确定为针对胎儿组织(如胎盘)的ncRNAs是microRNAs(MiRNAs)组的成员。可以预期的是，相当大的其他ncRNA群体会发挥更强大的影响和作用。因此，我们认为有必要扩大关于胎盘中的ncRNAs和从胎盘中释放的ncRNAs的知识。在胎盘内，胎儿的合体滋养层细胞形成胎儿和母亲之间的界面，外体和微囊从那里永久地释放到母亲的血液循环中。这些颗粒含有胎儿蛋白质和RNA，用于与邻近和远处的母体细胞进行通讯。颗粒的数量、大小和含量可以反映或预测胎盘紊乱，并可以在孕妇血清样本中进行评估。由于许多已知的滋养层细胞特异性miRNAs是人类灵长类动物所特有的，因此可以预期，其他目前未知的ncRNAs也是物种受限的，因此只能在人类胎盘中进行研究。一些严重的妊娠病理，包括先兆子痫，是人类特有的，它们的病理机制尚不清楚。我们期望关于新的胎盘ncRNA的知识将有可能彻底改变对胎盘内部调控过程和胎儿与母体沟通的理解。在这个拟议的项目中，我们旨在回答以下问题：(1)哪些(部分新鉴定的)ncRNA是胎儿特有的？(2)这些在胎儿中表达的ncRNA中哪些来自滋养层细胞？哪些滋养细胞来源的ncRNA到达母体血液中，包裹在(3)微泡或(4)外体中？此外，我们的目标是通过从母体血清中通过聚合酶链式反应鉴定高度丰富的ncRNAs。这可能有助于在后续项目中识别与疾病相关的标记基因。最后，我们计划确定自然滋养层细胞和常用细胞系(JEG-3绒毛膜癌细胞和HTR8/SVneo永生化早期滋养层细胞)中ncRNAs的差异。随后，两种细胞系都将被用于沉默和过度表达滋养层细胞特异性的ncRNA，以确定其靶点和基本功能。

Non-coding RNAs (ncRNAs) play a central and regulatory role in almost all cells, organs, and species which has been broadly recognized since the human ENCODE project and several other genome projects. Nevertheless, we know only a small fraction of ncRNAs. Moreover, long non-coding RNAs are known only for about five years and have been very marginally investigated in the placenta. To date, most examples of ncRNAs which have been identified to be specific for fetal tissues, such as placenta, are members of the group of microRNAs (miRNAs). It can be expected that the fairly larger group of other ncRNA exerts much more powerful influences and effects. Thus, we see the need to expand the knowledge about ncRNAs in the placenta and ncRNAs released from it.Inside the placenta, the fetal syncytiotrophoblast forms the interface between fetus and mother, from which exosomes and microvesicles are permanently released into the maternal circulation. These particles contain fetal proteins and RNA for communication with neighboring and distant maternal cells. The number, size and content of particles may reflect or predict placental disorders and can be assessed in maternal serum samples. As many of the known trophoblast-specific miRNAs are human-primate-specific it may be expected that also other, currently unknown ncRNAs are species-restricted and can, therefore, only be investigated in human placentae. Several severe pregnancy pathologies, including preeclampsia, are human-specific and their pathomechanisms are not yet understood. We expect that the knowledge on novel placental ncRNA will have the potential of revolutionizing the understanding of regulation processes inside the placenta and of fetal-maternal communication.In this proposed project, we aim to answer the following questions:(1) Which (partially newly characterized) ncRNAs are fetus specific?(2) Which of these fetally expressed ncRNAs are derived from trophoblast cells? Which trophoblast-derived ncRNAs reach maternal blood packed within (3) microvesicles or (4) exosomes? Furthermore, we aim to identify highly abundant ncRNAs via PCR from maternal serum. This may contribute to identification of disease associated marker genes in subsequent projects. Finally, we plan to determine differences between natural trophoblast ncRNAs and ncRNAs in commonly used cell lines (JEG-3 choriocarcinoma cells and HTR8/SVneo immortalized first trimester trophoblast cells). Subsequently, both cell lines will be used for silencing and overexpression of trophoblast specific ncRNA for identification of their targets and basic functions.