Neuronal excitability and copy number variation disorders

摘要 拷贝数变异(CNV)是神经发育障碍的主要原因，但它们的生物学 调查和药理学靶向带来了许多挑战。删除部分 轨迹是 其中 这个 多数 频繁 成因 的 自闭症 光谱 紊乱 和16p11.2的复制 (ASD)。然而，在变化中 相应的蛋白质网络，特别是在致病的关键细胞位置上，还没有被研究过 这个或其他CNV。我们建议使用隔室特定的神经蛋白质组学，结合生物信息学， 超分辨显微镜和药物再利用，以了解和改变树突状细胞的兴奋性表型 在16p11.2小鼠和诱导多能干细胞(IPSC)模型中。根据我们广泛的初步数据， 我们假设PRRT2的表达变化，它可能调节离子的子集的运输 血浆离子通道和受体的通道和受体、驱动力和异常补体 膜，导致异常的兴奋性，兴奋/抑制(E/I)平衡和网络特性 16p11.2模型和患者。这些表型可以通过使用FDA靶向离子通道功能来逆转。 批准的抗癫痫药物或使用重新调整用途的抗癌药物的ERK信号。我们的协作团队， 包括神经发育障碍(Penze)、神经蛋白质组学(SAVAS)、分子药理学方面的专家 (Barbolina)和离子通道生理学(George)将采用强大的多学科组合 高度创新的方法，以追求以下具体目标：(1)绘制发展图 调节和确定DUP和Del小鼠异常兴奋性的分子机制 人类神经元。(2)绘制发育概况图并确定潜在的分子机制 PRRT2作为兴奋性和癫痫表型的驱动因素的作用。(3)16p11.2 del的药理逆转 和DUP表型。这项提案将第一次证明细胞亚舱特有 蛋白质组学与超分辨显微镜相结合，由高渗透性单基因疾病提供信息 CNV中的基因，可以识别新的疾病机制。这种表型可以通过以下方式在全球范围内逆转 使用重新调整用途的药物靶向网络中心，开启治疗高血压的新策略 神经发育障碍。

ABSTRACT Copy number variations (CNVs) are a major cause of neurodevelopmental disorders, but their biological investigation and pharmacological targeting pose many challenges. Deletions locus are among the most frequent causes of autism spectrum disorder and duplications at the 16p11.2 (ASD). However, alterations in the corresponding protein networks, especially at key cellular sites for pathogenesis, have not been investigated in this or other CNVs. We propose to use compartment-specific neuroproteomics, combined with bioinformatics, super-resolution microscopy, and drug repurposing, to understand and alter dendritic excitability phenotypes in 16p11.2 mouse and induced pluripotent stem cell (iPSC) models. Based on our extensive preliminary data, we hypothesize that altered expression of PRRT2, which likely regulates the trafficking of a subset of ion channels and receptors, drives and abnormal complement of ion channels and receptor on the plasma membrane, leading to abnormal excitability, excitatory/inhibitory (E/I) balance, and network properties in 16p11.2 models and patients. These phenotypes may be reversed by targeting ion channel function using FDA- approved anti-epileptic drugs or ERK signaling using repurposed cancer drugs. Our collaborative team, which includes experts in neurodevelopmental disorders (Penzes), neuroproteomics (Savas), molecular pharmacology (Barbolina), and ion channel physiology (George) will employ a powerful and multidisciplinary combination of highly innovative methodologies to pursue the following Specific Aims: (1) To chart the developmental regulation and determine molecular mechanisms underlying abnormal excitability in dup and del mice and human neurons. (2) To chart the developmental profile and determine the molecular mechanisms underlying the role of PRRT2 as a driver of excitability and seizure phenotypes. (3) Pharmacological reversal of 16p11.2 del and dup phenotypes. This proposal will be the first to demonstrate that cellular subcompartment-specific proteomics combined with super-resolution microscopy, informed by highly penetrant monogenic disease genes within a CNV, can identify novel disease mechanisms. Such phenotypes could be reversed globally by targeting network hubs using repurposed drugs, opening novel strategies for the treatment of neurodevelopmental disorders.