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，但其生物学特性可能与神经发育障碍有关。 研究和药理学靶向提出了许多挑战。缺失 基因座 之间 的 最 频繁 原因 的 自闭症 频谱 障碍 16p11.2的重复 （ASD）。然而， 相应的蛋白质网络，特别是在发病机制的关键细胞位点，还没有研究， 或者其他的CNV。我们建议使用区室特异性神经蛋白质组学，结合生物信息学， 超分辨率显微镜和药物再利用，以了解和改变树突兴奋性表型 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PRRT 2的表达改变，可能调节一个亚类离子的运输， 离子通道和受体，血浆上离子通道和受体的驱动和异常补体 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ERK信号转导使用再利用的癌症药物。我们的合作团队， 包括神经发育障碍（Penzes）、神经蛋白质组学（Savas）、分子药理学 （Barbolina）和离子通道生理学（乔治）将采用一个强大的和多学科的组合， 高度创新的方法，以追求以下具体目标：（1）制定发展 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)绘制发育概况并确定其分子机制 PRRT 2作为兴奋性和癫痫发作表型驱动因素的作用。(3)16p11.2缺失的药理学逆转 和DUP表型。该提案将首次证明细胞亚区室特异性 蛋白质组学与超分辨率显微镜相结合，由高度渗透的单基因疾病提供信息 CNV内的基因，可以识别新的疾病机制。这种表型可以在全球范围内逆转， 使用重新用途的药物瞄准网络中心，为治疗 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.