Examining post-transcriptional regulation of the segmentation clock by microRNAs

检查 microRNA 对分段时钟的转录后调节

• 批准号: 8053365
• 负责人: SUSAN E COLE
• 金额: $ 7.21万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2013-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8053365
• 关键词: 3' Untranslated Regions; Binding; Biological Assay; Cell Culture Techniques; Congenital Abnormality; Coupled; Data; Defect; Disease; Dysostoses; Embryo; Embryonic Development; Exhibits; FOLH1 gene; Feedback; Future; Gene Expression; Gene Targeting; Genes; Genetic; Genetic Transcription; Genetic Translation; Half-Life; Hour; In Situ Hybridization; Link; Messenger RNA; Methods; MicroRNAs; Mission; Mus; Mutation; National Institute of Child Health and Human Development; Nucleotides; Pathway interactions; Pattern; Play; Post-Transcriptional Regulation; Process; Production; Public Health; RNA; RNA Binding; RNA Stability; Regulation; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Segmentation Clock Pathway; Signal Pathway; Skeletal Development; Skeleton; Somites; Techniques; Testing; Time; Transcript; Transcriptional Regulation; Untranslated Regions; Vertebral column; base; candidate validation; falls; interest; mRNA Expression; mRNA Stability; member; notch protein; programs; research study; rib bone structure; somitogenesis

DESCRIPTION (Provided by Applicant): The vertebrate embryonic axis consists of serially repeated lements, the most evident of which are the somites. The proper production of somites during somitogenesis is critical to the formation of the axial skeleton. The process of somitogenesis is regulated in part by a genetic clock. Key genes in the Notch, Wnt, and Fgf pathways exhibit cyclic expression with a period that matches the rate of somite formation. Mutations that perturb the proper oscillation of these genes can result in congenital defects of the axial skeleton, including spondylocostal dysostoses. Thus, understanding the regulatory mechanisms that control segmentation clock function is critical to our understanding of skeletal development. Several groups have examined the transcriptional control of oscillatory gene expression linked to the segmentation clock, but comparatively little is known about the post-transcriptional mechanisms that regulate clock function. The investigator hypothesizes that microRNAs may play important roles in the post-transcriptional control of genes linked to the clock. miRNAs are small conserved RNAs that bind to target transcripts and promote mRNA turnover and/or inhibit mRNA translation. She proposes that the function of these miRNAs may contribute to the delays in the negative feedback regulatory loops that underlie oscillatory RNA expression. She has identified miRNAs that are enriched in a pattern that is consistent with potential functions in the segmentation clock, and find that many of these miRNAs are predicted to target the 3'UTRs of oscillatory genes. Identifying miRNAs that function in the segmentation clock would provide a new paradigm to understand the posttranscriptional regulation of the segmentation clock. To pursue this idea it will be critical to generate further preliminary data connecting these candidate miRNAs to potential functions in the clock. The investigator therefore proposes to examine these miRNAs during somitogenesis by validating the enrichment of these candidate miRNAs, examining their expression patterns throughout embryogenesis, and functionally testing their ability to target and regulate expression of oscillatory genes. The experiments proposed here fall directly in the scope of the R03 program, as they represent a small, self-contained project that will produce critical preliminary data for future research. Further, this research falls directly into the mission of the NICHD, examining the mechanisms by which the spatial and temporal control of gene expression during embryogenesis functions to regulate somitogenesis. It is predicted that the research proposed here will provide critical links between miRNA regulation and segmentation clock function, and will provide a base of preliminary data that will support further research into the actual functions of miRNAs during somitogenesis as well as the segmentation clock in a future R01 application. RELEVANCE: The processes of embryonic development that produce the spine and ribs are controlled by a genetic clock. When this clock is perturbed, the results can be congenital defects. This research examines different mechanisms that control the expression of genes that have been linked to this clock. By understanding how the expression of genes in the clock is controlled, we will be better able to target treatments for defects and diseases that arise from misregulation of the clock.

描述(申请人提供)：脊椎动物胚轴由连续重复的元素组成，其中最明显的是体节。在体细胞发生过程中，体节的适当产生对中轴骨骼的形成至关重要。体细胞发生的过程在一定程度上受遗传时钟的调节。Notch、Wnt和Fgf途径中的关键基因表现出周期性的表达，其周期与体节形成的速率相匹配。扰乱这些基因正常振荡的突变可能会导致中轴骨的先天性缺陷，包括脊柱肋骨发育不良。因此，了解控制分段时钟功能的调控机制对于我们理解骨骼发育至关重要。 几个小组已经研究了与分段时钟相关的振荡基因表达的转录控制，但对调控时钟功能的转录后机制知之甚少。研究人员假设，microRNAs可能在与时钟相关的基因的转录后控制中发挥重要作用。MiRNAs是一种小的、保守的RNA，它与靶转录产物结合，促进mRNA的周转和/或抑制mRNA的翻译。她提出，这些miRNAs的功能可能导致了作为RNA振荡表达基础的负反馈调控环路的延迟。她已经确定了以与分段时钟中潜在功能一致的模式富含的miRNAs，并发现其中许多miRNAs被预测以振荡基因的3‘UTRs为靶标。识别在分段时钟中起作用的miRNAs将为理解分段时钟的转录后调控提供一个新的范式。为了实现这一想法，关键是产生更多的初步数据，将这些候选miRNAs与时钟中的潜在功能联系起来。 因此，研究人员建议通过验证这些候选miRNAs的浓缩，检测它们在整个胚胎发育过程中的表达模式，并从功能上测试它们靶向和调节振荡基因表达的能力，来检测这些miRNAs在体细胞发生过程中的作用。这里提出的实验直接属于R03计划的范围，因为它们代表了一个小型的、独立的项目，将为未来的研究提供关键的初步数据。此外，这项研究直接属于NICHD的任务，研究胚胎发育过程中基因表达的空间和时间控制功能调节体细胞发育的机制。可以预见，本研究将提供miRNA调控与分段时钟功能之间的关键联系，并将为进一步研究miRNAs在体细胞发生中的实际功能以及未来R01应用中的分段时钟提供初步数据基础。 相关性：产生脊椎和肋骨的胚胎发育过程受基因时钟控制。当这个时钟被扰乱时，结果可能是先天缺陷。这项研究考察了控制与生物钟相关的基因表达的不同机制。通过了解时钟中基因的表达是如何控制的，我们将能够更好地针对由于时钟调节不当而引起的缺陷和疾病进行治疗。

项目成果

Mir-125a-5p-mediated regulation of Lfng is essential for the avian segmentation clock.

• DOI: 10.1016/j.devcel.2013.01.024
• 发表时间: 2013-03-11
• 期刊: DEVELOPMENTAL CELL
• 影响因子: 11.8
• 作者: Riley, Maurisa F.;Bochter, Matthew S.;Wahi, Kanu;Nuovo, Gerard J.;Cole, Susan E.
• 通讯作者: Cole, Susan E.