Dynamic control of cortical development and disease by mRNA stability

摘要 大脑皮层的发育需要精确的调节控制，以确保基因在 正确的细胞类型和适当的发育阶段。跨细胞类型和基因表达的分析 发展，揭示了细胞转录本的广泛多样性。然而，仅转录数据集 提供基因表达的静态快照。转录本的表达受到不同速率的影响 转录和RNA降解，但后者在皮质发育中的作用鲜为人知。这个 这项提议的总体目标是揭示信使核糖核酸的稳定性如何有助于细胞的动态性质 大脑皮层发育的转录本。首先，我们将第一次阐明，时间景观 大脑皮质发育过程中信使核糖核酸的稳定性我们将使用最先进的SLAM-SEQ方法来量化 小鼠和人类模型中神经分化阶段的信使核糖核酸稳定性。这些数据将是 询问以告知顺式和反式控制RNA降解并确定受以下因素影响的疾病位点的优先顺序 稳定性。其次，我们将调查对CCR4组件的要求-不是复杂的，中央监管机构 在大脑皮层发育过程中，RNA的稳定性。我们的初步数据表明，CNOT3组件是必不可少的 用于大脑皮层发育。因此，我们将使用击倒方法和救援实验来衡量 作为CCR4的一部分，CNOT3控制皮质发育的程度--不复杂。更进一步，如 在自闭症中，这种复合体的成分发生突变，我们将研究这些突变是如何损害大脑皮层的 发展。完成后，我们的研究将建立第一个全面的mRNAs库，这些mRNAs是 在老鼠和人类发育的稳定水平上进行动态调节。通过研究基因 RNA稳定性因子对皮质生成的要求，我们将建立细胞命运的新范式 在神经系统中受到调节，与疾病相关。总之，这将开辟新的研究方向。 进入大脑发育和疾病的分子控制的基础。

Abstract Development of the cerebral cortex requires precise regulatory control to ensure genes are expressed in the correct cell types and at the proper developmental stages. Analyses of gene expression across cell types and development, have revealed extensive diversity in cellular transcriptomes. However, transcriptomic datasets only provide a static snapshot of gene expression. Transcript expression is influenced by a balance between rates of transcription and RNA degradation, yet the role of the latter in cortical development is poorly understood. The overall goal of this proposal is uncover how mRNA stability contributes to the dynamic nature of cellular transcriptomes across cortical development. First, we will elucidate, for the first time, the temporal landscape of mRNA stability during cortical development. We will use state-of-the-art SLAM-seq approaches to quantify mRNA stability across neural differentiation stages in both mouse and human models. These data will be interrogated to inform both cis and trans control of RNA degradation and to prioritize disease loci influenced by stability. Second, we will investigate requirements for components of the CCR4-NOT complex, a central regulator of RNA stability, in cortical development. Our preliminary data demonstrate the CNOT3 component is essential for cortical development. Thus, we will use knockdown approaches and rescue experiments to measure the extent to which CNOT3 controls cortical development as part of the CCR4-NOT complex. Further, as components of this complex are mutated in autism, we will investigate how these mutations impair cortical development. Upon completion, our studies will establish the first comprehensive repertoire of mRNAs that are dynamically regulated at the level of stability in mouse and human development. By investigating genetic requirements of RNA stability factors for corticogenesis, we will establish new paradigms for how cell fate is regulated in the nervous system and relevant for disease. Together, this will open up new research directions into the basis of molecular control of brain development and disease.