Post-transcriptional RNA regulation in mammalian neural stem cells

哺乳动物神经干细胞的转录后RNA调控

基本信息

批准号：
9317830
负责人：
Debra Silver
金额：
$ 19.88万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-04-01 至 2019-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9317830
关键词：
1q21 Address Affect Antibodies Attention Autistic Disorder Binding Biochemical Bone Diseases Brain Catalogs Cell Line Cells Cerebral cortex Complex Copy Number Polymorphism Data Data Set Development Diagnostic Disease Embryo Etiology Exons Foundations Future Genes Genetic Genetic Transcription Goals Hematological Disease Human Immunoprecipitation Intellectual functioning disability Investigation Knowledge Macrocephaly Measures Mediating Mental disorders Messenger RNA Metabolism Microcephaly Modeling Molecular Molecular Target Mus Mutation Neurodevelopmental Deficit Neurodevelopmental Disorder Neuroepithelial Neurons Pathologic Pathology Pathway interactions Phenotype Polyribosomes Post-Transcriptional Regulation Process Production Proliferating Proteins Proteomics RNA RNA Splicing RNA immunoprecipitation sequencing RNA-Binding Proteins Regulation Role Signal Pathway Sum Syndrome Testing Therapeutic Transcript Translational Regulation Translations base brain size deep sequencing developmental disease immortalized cell in vivo insight microdeletion mutant nerve stem cell nervous system disorder neurogenesis neuroregulation novel novel strategies progenitor self-renewal stem transcriptome

项目摘要

ABSTRACT This goal of this R21 proposal is to uncover post-transcriptional mechanisms important for neural progenitors of the developing brain, by focusing on the RNA binding protein, Rbm8a. RBM8A copy number variations are associated with intellectual disability, autism, microcephaly and macrocephaly. RBM8A mutations cause TAR syndrome, a rare blood and bone disorder which can also present with neurodevelopmental deficits. These neurodevelopmental pathologies can arise from aberrant neurogenesis of the cerebral cortex, in which neurons are produced from proliferating neural progenitors. Indeed, our lab has discovered that neural progenitor- specific Rbm8a haploinsufficiency in mice disrupts progenitor function and neurogenesis, which ultimately causes microcephaly. Rbm8a, together with Magoh and Eif4a3, form the exon junction complex, which binds spliced mRNAs to mediate key aspects of mRNA metabolism including translation. Together these findings demonstrate Rbm8a may influence disease by disrupting neurogenesis. However, our understanding of molecular mechanisms by which Rbm8a impacts brain development is limited to just a few candidates. In this proposal, we will test the central hypothesis that Rbm8a binds to and promotes translation of transcripts relevant for neural progenitor proliferation. First we will identify direct targets of Rbm8a in neuroepithelial progenitors, using a novel RNA immunoprecipitation-based approach in vivo. Second, we will use biochemical approaches to discover Rbm8a translational targets in neuroepithelial progenitors. Our proposal is built upon strong preliminary data, including the discovery of transcriptome-wide changes in Rbm8a hapoloinsufficient neuroepithelial progenitors. Successfully completed, these aims will provide new insights into how neural progenitors regulate fate decisions in the developing brain under both normal and pathological settings. Because RBM8A mutation is associated with neurodevelopmental disorders, these studies promise to provide important molecular insights into how mutations in this gene influence disease pathology.

抽象的 R21 提案的目标是揭示对神经祖细胞重要的转录后机制通过关注 RNA 结合蛋白 Rbm8a 来观察发育中的大脑。 RBM8A 拷贝数变异是与智力障碍、自闭症、小头畸形和大头畸形有关。 RBM8A突变导致TAR 综合征，一种罕见的血液和骨骼疾病，也可能出现神经发育缺陷。这些神经发育病理学可由大脑皮层的异常神经发生引起，其中神经元由增殖的神经祖细胞产生。事实上，我们的实验室发现神经祖细胞小鼠中特定的 Rbm8a 单倍体不足会破坏祖细胞功能和神经发生，最终导致小头畸形。 Rbm8a 与 Magoh 和 Eif4a3 一起形成外显子连接复合物，该复合物结合剪接 mRNA 介导 mRNA 代谢的关键方面，包括翻译。综合这些发现证明 Rbm8a 可能通过破坏神经发生来影响疾病。然而，我们的理解 Rbm8a 影响大脑发育的分子机制仅限于少数候选者。在这个提案中，我们将测试 Rbm8a 结合并促进转录本翻译的中心假设与神经祖细胞增殖有关。首先，我们将确定神经上皮中 Rbm8a 的直接靶标祖细胞，使用一种新型的基于 RNA 免疫沉淀的体内方法。其次，我们将使用生化发现神经上皮祖细胞中 Rbm8a 翻译靶点的方法。我们的建议是建立在强有力的初步数据，包括 Rbm8a 单倍体转录组范围变化的发现不足神经上皮祖细胞。如果成功完成，这些目标将为神经网络如何发挥作用提供新的见解。祖细胞在正常和病理环境下调节发育中大脑的命运决定。由于 RBM8A 突变与神经发育障碍相关，因此这些研究有望提供关于该基因突变如何影响疾病病理学的重要分子见解。