Control of neuron activity and animal behavior by non-coding RNAs

非编码 RNA 控制神经元活动和动物行为

• 批准号: 9319339
• 负责人: Anne Schaefer
• 金额: $ 36.94万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-09-01 至 2019-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9319339
• 关键词: Address; Affect; Animal Behavior; Antiepileptic Agents; Attenuated; Behavior Control; Behavioral; Brain; CRISPR/Cas technology; Calcineurin; Calcium; Cell physiology; Chemicals; Collaborations; Data; Dependovirus; Development; Disease; Electrophysiology (science); Environmental Risk Factor; Epilepsy; Feedback; Gene Delivery; Gene Expression; Gene Mutation; Genes; Genetic; Genetic Transcription; Human; Hyperactive behavior; Laboratories; Link; Mediating; Mental disorders; Messenger RNA; Metabolism; MicroRNAs; Molecular; Mus; Mutagenesis; Mutant Strains Mice; Neurons; Pathway interactions; Pharmaceutical Preparations; Play; Porifera; Process; Prosencephalon; Proteins; RNA Binding; Regulation; Research; Research Design; Resistance; Role; Seizures; Signal Pathway; Signal Transduction; Signaling Protein; Syndrome; Technology; Testing; Therapeutic; Therapeutic Intervention; Translations; Untranslated RNA; base; crosslinking and immunoprecipitation sequencing; dosage; embryonic stem cell; experimental study; functional outcomes; in vivo; innovation; link protein; mRNA Expression; mouse model; nervous system disorder; neuron development; neurotransmission; novel; overexpression; pointed protein; postnatal; public health relevance; response; ribosome profiling

 DESCRIPTION (provided by applicant): The current project addresses a novel regulatory mechanism of neuronal activity by microRNAs and its role in controlling animal behavior and epilepsy. We found that a neuron-enriched microRNA, miR-128, acts as a negative master modulator of neuronal signaling responses in mice. The importance of the miR-128 in brain function is underscored by the development of fatal epileptic seizures in mice that lack miR-128 in postnatal forebrain neurons. Accordingly, overexpression of miR-128 in neurons attenuates chemically-induced seizures and rescues survival in mice. These findings reveal miR-128 as a previously unknown key regulator of neuronal activity with a potential for the development of novel anti-epileptic therapeutic approaches. The major regulatory function of miR-128 highlights the importance of the mechanisms that control miR-128 expression levels as well as miR-128 access to its targets in neurons. We found that calcineurin/NFAT activity, an important signaling pathway in neuronal development and function, controls miR-128 expression in neurons. Most importantly, mice with neuronal calcineurin deficiency develop a fatal seizure syndrome similar to the one observed in mice with miR-128 deficiency. These findings provided a strong indication for the role of calcineurin/NFAT signaling in control of miR-128 expression. The mechanism of calcineurin/NFAT-mediated regulation of miR-128 expression will be addressed in this proposal. The proposal also addresses a novel mechanism of regulation of miR-128 effector function. Preliminary data suggest that access of miR-128 to its mRNA targets is regulated by miR-128-sequestring, non-coding decoy RNAs. This novel mechanism of regulation of miR-128 effector function will be explored in experiments using unique mouse models with inactivation or overexpression of the neuron-specific miR-128 decoy RNAs in combination with the molecular, electrophysiological and behavioral approaches that have been established in our laboratory. The central role of miR-128 in neuronal signaling makes this miRNA an attractive target for potential therapeutic intervention of epilepsy. Using a mouse model of the human Dravet syndrome, one the most severe and often treatment resistant forms of epilepsy, the proposal will explore the therapeutic potential of neuronal miR-128 modulation for the treatment of the disease. To bring the research closer to the development of actual antiepileptic therapy, we will explore the antiepileptic potential of exogenously expressed miR-128. MiR-128 will be delivered to defined neurons using neurotrophic adeno-associated viruses (AAV) that are currently considered as the most reliable vehicle for in vivo gene delivery in mice and human. In summary, our proposal describes highly innovative and hypothesis driven studies of a novel non-coding RNA mechanism for controlling neuronal signaling and activity that has a strong potential to be applied for the treatment of human epilepsy.

 描述（由适用提供）：当前的项目解决了microRNA的神经元活动的新调节机制及其在控制动物行为和癫痫中的作用。我们发现，富含神经元的microRNA miR-128充当小鼠神经元信号反应的负主调节剂。 miR-128在脑功能中的重要性被小鼠致命性癫痫发作的发展所缺乏，这些癫痫发作缺乏产后前脑神经元中缺乏miR-128的小鼠。根据上述，神经元中miR-128的过表达会减弱化学诱导的癫痫发作并挽救小鼠的存活率。这些发现揭示了miR-128是一种以前未知的神经元活性的关键调节剂，具有开发新型抗癫痫疗法方法的潜力。 miR-128的主要调节功能强调了控制miR-128表达水平以及miR-128在神经元中的靶标的机制的重要性。我们发现钙调神经酶/NFAT活性是神经元发育和功能中重要的信号通路，它控制着神经元中的miR-128表达。最重要的是，神经元钙调蛋白缺乏症的小鼠会形成一种致命的癫痫综合征，类似于在具有miR-128缺乏症的小鼠中观察到的小鼠。这些发现为钙调蛋白/NFAT信号在控制miR-128表达中的作用提供了有力的指示。该提案将解决钙调蛋白/NFAT介导的miR-128表达调节的机理。该提案还解决了miR-128效应函数调节的新型机制。初步数据表明，miR-128对其mRNA靶标的访问受miR-128序列的非编码诱饵RNA调节。在我们实验室中确定的分子，电生理学和行为方法结合使用，将在实验中探索这种新型MiR-128效应功能调节效应功能的机理。 miR-128在神经元信号传导中的核心作用使该miRNA成为癫痫潜在治疗干预的有吸引力的靶标。该提案使用人类Dravet综合征的小鼠模型是最严重且经常抗治疗的癫痫形式的一种，将探索神经元miR-128调节对疾病治疗的治疗潜力。为了使研究更接近实际抗癫痫疗法的发展，我们将探索极其表达的miR-128的抗癫痫潜力。 miR-128将使用神经营养相关病毒（AAV）传递到定义的神经元中，这些病毒目前被认为是小鼠和人体内基因递送最可靠的载体。总而言之，我们的建议描述了对控制神经元信号传导和活性的新型非编码RNA机制的高度创新和假设驱动的研究，该机制具有很强的潜力，可用于治疗人癫痫。