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

 描述（由申请人提供）：目前的项目解决了一个新的神经元活动的microRNA调控机制及其在控制动物行为和癫痫的作用。我们发现，一种富含神经元的microRNA，miR-128，在小鼠中作为神经元信号反应的负主调节剂。在出生后前脑神经元中缺乏miR-128的小鼠中，致命性癫痫发作的发展强调了miR-128在脑功能中的重要性。因此，神经元中miR-128的过表达减弱了化学诱导的癫痫发作并挽救了小鼠的存活。这些发现揭示了miR-128作为一种以前未知的神经元活性的关键调节因子，具有开发新型抗癫痫治疗方法的潜力。miR-128的主要调节功能突出了控制miR-128表达水平以及miR-128在神经元中接近其靶点的机制的重要性。我们发现，钙调磷酸酶/NFAT活性，神经元发育和功能的重要信号通路，控制神经元中的miR-128表达。最重要的是，神经元钙调磷酸酶缺乏的小鼠发生致命性癫痫发作综合征，与在miR-128缺乏的小鼠中观察到的相似。这些发现为钙调磷酸酶/NFAT信号传导在控制miR-128表达中的作用提供了强有力的指示。钙调神经磷酸酶/NFAT介导的miR-128表达调控机制将在本提案中得到解决。该提案还提出了一种调节miR-128效应子功能的新机制。初步数据表明，miR-128与其mRNA靶点的接近受miR-128-螯合物、非编码诱饵RNA的调节。这种新的调节miR-128效应器功能的机制将在实验中探索，使用独特的小鼠模型与神经元特异性miR-128诱饵RNA的失活或过表达结合分子，电生理和行为方法，已经建立在我们的实验室。miR-128在神经元信号传导中的核心作用使该miRNA成为潜在的癫痫治疗干预的有吸引力的靶点。使用人类Dravet综合征的小鼠模型，这是最严重且通常是治疗抵抗性的癫痫形式之一，该提案将探索神经元miR-128调节用于治疗该疾病的治疗潜力。为了使研究更接近实际抗癫痫治疗的发展，我们将探索外源性表达的miR-128的抗癫痫潜力。MiR-128将使用神经营养腺相关病毒（AAV）递送至限定的神经元，所述神经营养腺相关病毒（AAV）目前被认为是用于小鼠和人类体内基因递送的最可靠的载体。总之，我们的提案描述了一种新型非编码RNA机制的高度创新和假设驱动的研究，该机制用于控制神经元信号传导和活性，具有很强的应用于治疗人类癫痫的潜力。

 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.