Post-transcriptional Regulation of Trophoblast Differentiation

项目摘要(见说明)；项目三 着床是胚胎滋养外胚层和母体子宫内膜相互作用的过程，对于许多夫妇来说，这仍然是成功妊娠的根本障碍。然而，人们对调节这两种组织的分子机制知之甚少。MicroRNAs为理解这些机制提供了一种手段。一个单独的microRNA可以抑制数百个信使RNA，并且mlRNA-mRNA目标对已经被进化地选择来具有特定的细胞结果。 因此，发现调控特定细胞命运的miRNAs，然后对其下游靶标进行全面分析，为了解调控这些细胞命运的分子网络提供了一个窗口。此外，了解miRNAs在疾病中是如何被错误调控的，可以深入了解miRNAs的功能障碍 这些网络导致了异常的发展。这项资助建议使用miRNAs来剖析调控滋养层干细胞增殖和分化的机制，并确定这些机制在子痫前期是否以及如何被破坏，子痫前期是一种与滋养层分化缺陷相关的疾病。中心假设是，miRNA功能分析与系统的靶标识别和功能分析相结合，将揭示调节胎盘生长和 发展。这一假说得到了初步结果的支持，这些结果显示，小鼠滋养层干细胞(TSC)自我更新和分化的缺陷与全球miRNA丢失有关，这些缺陷可以通过重新引入单个miRNAs来挽救。我们将追求三个目标。首先，影响TSC自我更新和分化的miRNAs将通过全球敲除表型的救援筛选来确定， 侧写和击倒实验。第二，基因组和高通量功能分析将揭示对TSC分化充分和/或必要的miRNAs的mRNA靶点。 第三，早期的发现将扩展到人类滋养层祖细胞，重点放在先兆子痫患者中那些调控错误的miRNAs上。这种方法是创新的，它结合了miRNA功能筛选、目标识别和在鲜为人知的组织类型中的途径发现。这一研究意义重大，因为它将为理解胚胎外组织中的缺陷提供分子框架。 与植入和妊娠疾病有关。这项提议是一个更大的U54中心拨款的一部分，通过招募其他PI在滋养层子宫内膜相互作用、转录后调控和基因组/生物信息学方法方面的专业知识，拟议的工作将从该中心获得极大的好处。

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.