Novel regulation and function of the lncRNA Gomafu in human neurons

最近的研究发现了一个快速增长的长非编码 RNA (lncRNA) 列表，其中包含超过 200个核苷酸没有开放阅读框，但在调节基因表达中发挥关键作用，从而控制 神经干细胞维持、神经发生、神经元网络组装和突触可塑性。长链非编码RNA 在进化过程中，人类的转录组显着扩展，并且在大脑中表达最丰富。 人类 lncRNA 的复杂性被认为是认知进化的主要架构师的基础，但也 引入各种脑部疾病的脆弱性。事实上，在自闭症中观察到了 lncRNA 失调， 智力障碍、癫痫、神经退行性疾病和神经精神疾病，表明 lncRNA 失调导致各种脑部疾病的发病机制。然而，我们目前的 对人类神经元中 lncRNA 的调控和功能的了解仍处于起步阶段。 lncRNA Gomafu，最初被鉴定为与心肌梗塞相关的转录本，因此命名为 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 此外，Gomafu 基因敲除小鼠表现出类似焦虑的行为。在源自人类诱导的神经元中 多能干细胞 (hiPSC)，Gomafu 调节初级转录物的选择性剪接 (AS) 神经元发育和突触功能。重要的是，在皮质灰质中检测到 Gomafu 失调 来自精神分裂症患者死后大脑的物质和中间神经元。这些发现 共同表明 Gomafu 在控制正常大脑功能方面发挥着重要作用。然而，分子 调节人类 Gomafu 表达的机制仍有待探索。 Gomafu 如何失调 脑部疾病尚不清楚。此外，Gomafu 如何控制人类神经元转录组的 AS 仍然难以捉摸。 Gomafu 缺乏如何影响人类神经元发育尚不清楚。这个提议 攻击这些重要问题，旨在 1）描述调节的分子机制和途径 hiPSC 衍生神经元中的 Gomafu 表达，尤其是遗传-表观遗传相互作用网络 围绕我们的初步数据揭示的新型 microRNA-lncRNA 功能相互作用； 2）确定 通过深度 RNA 测序确定人神经元转录组中 Gomafu 的选择性剪接靶点； 3） 确定 Gomafu 在 hiPSC 衍生的皮质兴奋性神经元发育中的功能 2-D 培养物和 3-D 类器官中的多巴胺能神经元。这些问题的答案将填补主流 关于lncRNA如何控制人类神经元和lncRNA正常发育的知识差距 大脑功能障碍。

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.