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）的名单，这些RNA含有大于 没有开放阅读框但在调节基因表达中起关键作用的200个核苷酸， 神经干细胞维持、神经发生、神经元网络组装和突触可塑性。lncRNA 转录组在进化过程中在人类中显著扩增，并且在大脑中表达最丰富。 人类lncRNAs的复杂性被认为是认知进化的主要架构师，但也 为各种脑部疾病引入了弱点。事实上，在自闭症中观察到lncRNA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失调有助于各种脑部疾病的发病机制。然而，我们目前 对人类神经元中lncRNA的调节和功能的理解仍处于起步阶段。 lncRNA Gomafu是一种最初被确定与心肌梗死相关的转录本，因此被命名为 最近发现MIAT在大脑中含量最丰富，并与正常神经元发育有关 和认知状况。Gomafu在突触刺激和恐惧条件反射后迅速下调。在 此外，Gomafu敲除小鼠表现出焦虑样行为。在人类诱导的神经元中- 在多能干细胞（hiPSC）中，Gomafu调节对多能干细胞（hiPSC）的生长至关重要的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，Gomafu失调在皮质灰质中检测到， 精神分裂症患者死后脑组织中的物质和中间神经元。这些发现 共同表明，Gomafu在管理正常的大脑功能中起着重要作用。然而，分子 调节人Gomafu表达的机制仍然未被探索。Gomafu是如何在 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缺乏症如何影响人类神经元发育尚不清楚。这项建议 攻击这些重要的问题，旨在1）描绘分子机制和途径，调节 Gomafu在hiPSC衍生的神经元中的表达，特别是关于遗传-表观遗传相互作用网络 以我们初步数据揭示的一种新的microRNA-lncRNA功能相互作用为中心; 2）确定 通过深度RNA测序确定Gomafu在人类神经元转录组中的可变剪接靶点; 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普遍 关于lncRNA如何控制人类神经元的正常发育和lncRNA 大脑功能紊乱

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.