Novel Regulation and Function of the lncRNA Gomafu in Human Neurons

人类神经元中 lncRNA Gomafu 的新调控和功能

• 批准号: 10633129
• 负责人: Yue Feng
• 金额: $ 55.6万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-07-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10633129
• 关键词: 3-Dimensional; Affect; Alternative Splicing; Autopsy; Binding; Biological Markers; Biology; Brain; Brain Diseases; CRISPR/Cas technology; Cell Maintenance; Code; Cognition Disorders; Cognitive; Complex; Data; Development; Diagnosis; Disease; Epigenetic Process; Epilepsy; Etiology; Evolution; Gene Expression; Genes; Genetic; Genetic Transcription; Human; Impairment; Induced pluripotent stem cell derived neurons; Intellectual functioning disability; Interneurons; Knockout Mice; Knowledge; Mediating; Mental disorders; MicroRNAs; Midbrain structure; Molecular; Mus; Mutation; Myocardial Infarction; Names; Neurodegenerative Disorders; Neuronal Differentiation; Neurons; Nuclear; Nucleotides; Open Reading Frames; Organoids; Pathogenesis; Pathway interactions; Patients; Play; Pluripotent Stem Cells; Polypyrimidine Tract-Binding Protein; Prosencephalon; Proteins; RNA; Regulation; Reporting; Repression; Rodent; Role; Schizophrenia; Synapses; Synaptic plasticity; Testing; Therapeutic; Tissues; Transcript; Transcription Repressor; Untranslated RNA; anxiety-like behavior; autism spectrum disorder; cell type; conditioned fear; dopaminergic neuron; epigenetic regulation; excitatory neuron; gray matter; histone modification; infancy; knock-down; nerve stem cell; nervous system disorder; neural network; neurogenesis; neuron development; neuropsychiatric disorder; neuropsychiatry; novel; posttranscriptional; promoter; psychostimulant; risk variant; schizophrenia risk; synaptic function; synaptogenesis; transcription factor; transcriptome; transcriptome sequencing

Recent studies identified a fast growing list of long noncoding RNAs (lncRNAs) that harbor greater than 200 nucleotides with no open reading frames but play key roles in regulating gene expression thus govern neural stem cell maintenance, neurogenesis, neuronal network assembly, and synaptic plasticity. The lncRNA transcriptome is strikingly expanded in human during evolution and most abundantly expressed in the brain. The complexity of human lncRNAs is thought to underlie the major architect of cognitive evolution but also introduce vulnerabilities for various brain diseases. Indeed, lncRNA dysregulation is observed in autism, intellectual disability, epilepsy, neurodegenerative disorders and neuropsychiatric diseases, suggesting that lncRNA dysregulation contributes to the pathogenesis of various brain illnesses. However, our current understanding of regulation and function of lncRNAs in human neurons are still in the infancy. The lncRNA Gomafu, a transcript initially identified to associate with myocardial infarction thus named MIAT, was recently found to be most abundant in the brain and implicated in normal neuronal development and cognitive conditions. Gomafu is quickly downregulated upon synaptic stimulation and fear-conditioning. In addition, Gomafu knockout mice display anxiety-like behaviors. In neurons derived from human induced- pluripotent stem cells (hiPSCs), Gomafu regulates alternative splicing (AS) of primary transcripts essential for neuronal development and synaptic function. Importantly, Gomafu dysregulation is detected in cortical grey matters and interneurons of post-mortem brains derived from schizophrenia patients. These discoveries together suggest that Gomafu plays essential roles in governing normal brain function. However, molecular mechanisms that regulate human Gomafu expression remain unexplored. How Gomafu is dysregulated in brain diseases is not understood. Moreover, how Gomafu controls AS of the human neuronal transcriptome remains elusive. How Gomafu deficiency affects human neuron development is unknown. This proposal attacks these important questions, aiming to 1) Delineate molecular mechanisms and pathways that regulate Gomafu expression in hiPSC-derived neurons, especially regarding a genetic-epigenetic interaction network centering on a novel microRNA-lncRNA functional interplay revealed by our preliminary data; 2) determine the alternative splicing targets of Gomafu in human neuronal transcriptome by deep RNA-sequencing; 3) determine the function of Gomafu in the development of hiPSC-derived cortical excitatory neurons and dopaminergic neurons in 2-D culture and 3-D organoids. Answers to these questions will fill prevailing knowledge gaps regarding how lncRNAs govern normal development of human neurons and lncRNA malfunction in brain disorders.

最近的研究发现，越来越多的长非编码RNA(LncRNAs)含有超过 200个核苷酸没有开放阅读框，但在调节基因表达方面发挥关键作用，因此决定了 神经干细胞维持、神经发生、神经元网络组装和突触可塑性。印迹核糖核酸 转录组在人类进化过程中显著扩展，并在大脑中大量表达。 人类lncRNA的复杂性被认为是认知进化的主要基础，但也 引入各种脑部疾病的脆弱性。事实上，在自闭症患者中观察到了lncRNA调节失调， 智力残疾、癫痫、神经退行性疾病和神经精神疾病，这表明 LncRNA异常参与了多种脑部疾病的发病机制。然而，我们目前 对lncRNAs在人类神经元中的调节和功能的了解仍处于起步阶段。 LncRNA Gomafu，一种最初被认为与心肌梗死有关的转录本，因此被命名为 最近发现MIAT在大脑中含量最丰富，与正常神经元发育有关 和认知条件。Gomafu在突触刺激和恐惧条件作用下迅速下调。在……里面 此外，Gomafu基因敲除小鼠表现出类似焦虑的行为。在人类诱导产生的神经元中- 多能干细胞(HiPSCs)，Gomafu调节初级转录本的选择性剪接(AS) 神经元发育和突触功能。重要的是，Gomafu失调是在皮质灰质中检测到的 精神分裂症患者死后脑的物质和中间神经元。这些发现 综上所述，Gomafu在调节正常大脑功能方面发挥着至关重要的作用。然而，分子 调节人类Gomafu表达的机制仍未被探索。Gomafu是如何在 脑部疾病目前还不清楚。此外，Gomafu是如何控制人类神经元转录组的 仍然难以捉摸。Gomafu缺乏症如何影响人类神经元发育尚不清楚。这项建议 解决这些重要问题，目的是1)描绘调控的分子机制和途径 Gomafu在HiPSC来源神经元中的表达，特别是关于遗传-表观遗传相互作用网络 围绕我们的初步数据揭示的一种新的microRNA-lncRNA功能相互作用；2)确定 Gomafu在人神经元转录组中选择性剪接靶点的深度RNA测序；3) 确定Gomafu在HiPSC来源的皮质兴奋性神经元发育中的作用 2-D培养的多巴胺能神经元和3-D有机体。这些问题的答案将填满当前 关于lncRNA如何管理人类神经元和lncRNA正常发育的知识空白 脑部疾病的故障。