Post-transcriptional Regulation of Trophoblast Differentiation

项目概要（见说明）;项目III 着床是胚胎滋养外胚层与母体子宫内膜相互作用的过程，是许多夫妇成功妊娠的根本障碍。然而，对调节这两种组织的分子机制知之甚少。microRNA为理解这些机制提供了一种手段。单个microRNA可以抑制数百个信使RNA，并且mlRNA-mRNA靶对已经被进化选择为具有特定的细胞结果。 因此，揭示调节特定细胞命运的miRNA，然后对其下游靶标进行全面分析，为调节这些细胞命运的分子网络提供了一个窗口。此外，了解miRNAs如何在疾病中被错误调节，可以提供对功能障碍的深入了解。 这些网络导致了不正常的发展。该基金建议使用miRNAs来剖析滋养层干细胞增殖和分化的调节机制，并确定这些机制是否以及如何在先兆子痫中被破坏，先兆子痫是一种与滋养层分化缺陷相关的疾病。核心假设是，miRNA功能分析结合系统的靶点鉴定和功能分析将揭示调节胎盘生长的新途径， 发展这一假设得到了初步结果的支持，初步结果显示小鼠滋养层干细胞（TSC）自我更新和分化的缺陷与整体miRNA丢失相关，可以通过重新引入单个miRNA来挽救。将追求三个目标。首先，影响TSC自我更新和分化的miRNA将通过整体敲除表型的拯救筛选来确定， 侧写和击倒实验第二，通过基因组和高通量功能分析将揭示足够和/或必需用于TSC分化的miRNA的mRNA靶。 第三，早期的研究结果将扩展到人类滋养层祖细胞，重点是那些在先兆子痫患者中失调的miRNAs。该方法是创新的，它结合了miRNA功能筛选，靶点鉴定和在一个知之甚少的组织类型中的途径发现。这是重要的，因为它将提供分子框架，以了解缺陷的额外胚胎组织 与着床和妊娠相关的疾病。该提案是一个更大的U 54中心赠款的一部分，拟议的工作将大大受益于该中心通过招募其他Pis的滋养层子宫内膜相互作用，转录后调控，和基因组/生物信息学方法的专业知识。

PROJECT SUMMARY (See Instructions); PROJECT III Implantation, a process involving the interaction between the embryonic trophectoderm and maternal endometrium, remains a fundamental barrier to successful pregnancy for many couples. However, little is known about the molecular mechanisms that regulate either tissue. MicroRNAs provide a means to understand those mechanisms. An individual microRNA can suppress hundreds of messenger RNAs, and the mlRNA-mRNA target pairs have been evolutionarily selected to have specific cellular outcomes. Therefore, uncovering miRNAs that regulate specific cell fates followed by comprehensive analysis of their downstream targets provides a window into the molecular networks that regulate those cell fates. Furthermore, understanding how miRNAs are misregulated in disease can provide insight into how dysfunction of those networks results in aberrant development. This grant proposes to use miRNAs to dissect the mechanisms regulating trophoblast stem cell proliferation and differentiation and determine whether and how these mechanisms are disrupted in pre-eclampsia, a disease associated in defects of trophoblast differentiation. The central hypothesis is that miRNA functional analyses combined with systematic target identification and functional analyses will uncover novel pathways regulating placenta growth and development. This hypothesis is supported by preliminary results showing defects in mouse trophoblast stem cells (TSC) self-renewal and differentiation associated with global miRNA loss that can be rescued by re-introduction of individual miRNAs. Three aims will be pursued. First, miRNAs that influence TSC self renewal and differentiation will be determined through rescue screens of the global knockout phenotypes, profiling, and knockdown experiments. Second, mRNA targets of miRNAs that are sufficient and/or necessary for TSC differentiation will be uncovered by genomic and high throughput functional analyses. Third, early findings will be extended to human trophoblast progenitor cells focusing on those miRNAs misregulated in patients with pre-eclampsia. The approach is innovative in its combination of miRNA functional screens, target identification, and pathway discovery in a poorly understood tissue type. It is significant as it will provide the molecular framework to understand defects in extra embryonic tissues associated with diseases of implantation and pregnancy. The proposal is part of a larger U54 center grant The proposed work will benefit greatly from this center by recruiting the other Pis' expertise in trophoblast endometrium interactions, post-transcriptional regulation, and genomic/bioinformatic approaches.