Genetic analysis of a microRNA pathway regulating neural tube closure

调节神经管闭合的 microRNA 通路的遗传分析

• 批准号: 9075831
• 负责人: Jianfu Chen
• 金额: $ 32.81万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-04-01 至 2021-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9075831
• 关键词: 3' Untranslated Regions; Affect; Apoptotic; Behavior; Binding; Biological; Birth; Cell Proliferation; Cell Survival; Cell physiology; Chemicals; Complex; Congenital Abnormality; Cyclin D1; Data; Defect; Development; Embryo; Exhibits; Gene Expression; Gene Expression Regulation; Genes; Genetic; Goals; Human; Image; Individual; Knock-out; Lead; Mediating; Messenger RNA; Methods; MicroRNAs; Molecular; Morphogenesis; Mus; Mutant Strains Mice; Neural Tube Closure; Neural Tube Defects; Neural tube; Pathway interactions; Phenotype; Process; RNA-Binding Proteins; Regulator Genes; Repression; Role; Testing; Tissues; Untranslated RNA; base; cell behavior; embryo culture; genetic analysis; genetic approach; improved; inhibitor/antagonist; insight; loss of function; mouse model; mutant; neural plate; neural precursor cell; novel; premature; prevent; public health relevance; spatiotemporal; time use; trait

 DESCRIPTION (provided by applicant): The overall goal of this proposal is to establish a novel mechanism whereby microRNA (miRNA) miR-302/367 regulates neural tube closure (NTC). Disruption of NTC leads to neural tube defect (NTD), which is the second most common birth defect in humans affecting 1 to 1,000 births. However, the regulators and mechanisms that control NTC at post-transcriptional gene regulation levels remain largely unknown. We have created the first miRNA mouse model of NTD. We found that depletion of miR-302/367 leads to NTD and embryonic lethality. Neural precursor cells (NPCs) exhibit reduced proliferation, premature differentiation, and decreased survival in the mutant embryos. Importantly, we have identified individual miRNA targets with potent roles in specific cellular behaviors that are affected in mutant embryos. In addition, we found that miR-302/367 is associated with an RNA binding protein Lin41 to regulate gene expression; depletion of Lin41 also leads to NTD. These preliminary data lead to a novel hypothesis that miR-302/367 interacts with Lin41 to coordinately control multiple neural precursor cell (NPC) behaviors by regulating different miRNA targets during neural tube closure (NTC). In this project, we will determine miR-302/367 functions and their action mechanisms during neural tube closure. Three specific aims will be pursued: 1) Determine the developmental and cellular basis of neural tube defect (NTD) in miR-302/367 mutant mice; 2) Test whether individual genes that we have identified as targets of miR-302 mediate specific cellular behaviors during neural tube closure; 3) Test the hypothesis that miR-302 functions together with Lin41 to regulate gene expression and NPC behaviors during neural tube closure. Together, these studies will improve our understanding of genetic factors associated with NTD and provide novel insights into mechanisms underlying neural tube closure and neural tube defect (NTD).

 描述（由申请人提供）：本提案的总体目标是建立一种新的机制，通过 microRNA (miRNA) miR-302/367 调节神经管闭合 (NTC)。 NTC 破坏会导致神经管缺陷 (NTD)，这是人类第二常见的出生缺陷，影响 1 至 1,000 名新生儿。然而，在转录后基因调控水平上控制 NTC 的调控因子和机制仍然很大程度上未知。 我们创建了第一个 NTD miRNA 小鼠模型。我们发现 miR-302/367 的缺失会导致 NTD 和胚胎致死。神经前体细胞（NPC）在突变胚胎中表现出增殖减少、过早分化和存活率降低。重要的是，我们已经确定了在突变胚胎中受影响的特定细胞行为中具有有效作用的单个 miRNA 靶标。此外，我们发现miR-302/367与RNA结合蛋白Lin41相关以调节基因表达； Lin41 的耗尽也会导致 NTD。这些初步数据得出了一个新的假设，即 miR-302/367 与 Lin41 相互作用，通过在神经管闭合 (NTC) 过程中调节不同的 miRNA 靶标来协调控制多个神经前体细胞 (NPC) 行为。 在这个项目中，我们将确定 miR-302/367 在神经管闭合过程中的功能及其作用机制。我们将追求三个具体目标：1）确定 miR-302/367 突变小鼠神经管缺陷（NTD）的发育和细胞基础； 2) 测试我们确定为 miR-302 靶标的单个基因是否在神经管闭合过程中介导特定的细胞行为； 3) 检验 miR-302 与 Lin41 一起发挥作用以调节神经管闭合过程中的基因表达和 NPC 行为的假设。总之，这些研究将增进我们对与 NTD 相关的遗传因素的理解，并为神经管闭合和神经管缺陷 (NTD) 的潜在机制提供新的见解。