Targeting Mitochondrial Function to Develop Novel Therapies for Neurodevelopmental Disorders

摘要 我们将确定潜在的新的药物疗法，以改善线粒体的功能 被认为在多种神经发育障碍中受损的大脑皮层神经元。我们有 LgDel小鼠2/3层投射神经元线粒体代谢紊乱 22q11.2缺失综合征的模型，一种综合征性神经发育障碍。我们还展示了 这种干扰显然解释了大脑皮层连接性的数量差异 与LgDel小鼠的认知行为缺陷相关。最后，我们证明了一种自由基清除剂 影响线粒体功能的N-乙酰半胱氨酸(NAC)可以逆转这些分子、细胞和 行为缺陷。我们现在将评估多个线粒体靶向药理学的能力 化合物，以及已知的线粒体靶向药物，以调节LgDel层2/3PN线粒体 代谢及其对第2/3层PN动态平衡的影响。在具体目标1中，我们将评估化合物 使用安捷伦海马代谢分析仪进行验证的、均一的2/3层PN体外活性测定 测量线粒体功能的关键方面。修复LgDel线粒体的候选化合物 在不破坏WT线粒体功能的情况下，WT的功能障碍将进一步验证它们的能力 降低线粒体ROS水平，恢复树突状细胞和轴突 LgDel层2/3 PNS中的生长。提供对机械精度的其他解释性解决方案 这些化合物，在特定的目标2中，我们将进行LgDel和WT的平行转录组比较 Layer 2/3 PNS体外鉴定线粒体导致转录调控改变的途径 神经发育病理学所针对的发展中的第2/3层PNS的功能障碍和生长减少。 我们将进一步将这些数据与NAC处理的LgDEL 2/3层PNS的转录组图谱进行对比，NAC的 我们之前已经证明了代谢和恢复生长的活性。因此，在本文件中确定的化合物 将验证筛查的特异性、目标细胞活动，并将其置于不同监管的背景下 对线粒体功能改变敏感的转录途径。这些小路是被削弱的基础 有助于神经发育皮质回路病理的神经元生长。因此，我们的实验确定了 进一步开发基于线粒体的治疗和分子机制的潜在候选 用于纠正分子和细胞病理的精确靶向新药的合理设计框架 与多种神经发育障碍的皮质神经元和神经回路受损有关。

ABSTRACT We will identify potential new pharmacological therapies targeted to improve mitochondrial function in a class of cerebral cortical neurons thought to be compromised in multiple neurodevelopmental disorders. We have shown that mitochondrial metabolism is disrupted in layer 2/3 Projection Neurons (PNs) in the LgDel mouse model of 22q11.2 Deletion Syndrome, a syndromic neurodevelopmental disorder. We have also demonstrated that this disruption apparently accounts for quantitative differences in association cortical connectivity correlated with cognitive behavioral deficits in LgDel mice. Finally, we showed that a free radical scavenger that influences mitochondrial function, N-acetyl cysteine (NAC), can reverse these molecular, cellular and behavioral deficits. We will now assess the capacity of multiple mitochondrial targeted pharmacological compounds, as well as known mitochondria-targeted drugs, to modulate LgDel Layer 2/3 PN mitochondrial metabolism and its influences on layer 2/3 PN homeostasis. In Specific Aim 1, we will evaluate compound activity in a validated, homogeneous layer 2/3 PN in vitro assay using the Agilent Seahorse Metabolic Analyzer to measure key aspects of mitochondrial function. Candidate compounds that restore LgDel mitochondrial dysfunction toward WT, without disrupting WT mitochondrial function, will be further validated for their capacity to diminish aberrant mitochondrial reactive oxygen species (ROS) levels and restore dendritic and axonal growth in LgDel layer 2/3 PNs. To provide additional interpretative resolution of the mechanistic precision of these compounds, in Specific Aim 2 we will perform a parallel transcriptome comparison of LgDel versus WT Layer 2/3 PNs in vitro to identify pathways whose transcriptional regulation is altered due to mitochondrial dysfunction and diminished growth in developing Layer 2/3 PNs targeted by neurodevelopmental pathology. We will further contrast this data with the transcriptome profile of LgDel layer 2/3 PNs treated with NAC, whose metabolic and growth-restoring activity we have previously demonstrated. Thus, compounds identified in this screen will be validated for specificity, targeted cellular activity, and placed in context of differentially regulated transcriptional pathways sensitive to altered mitochondrial function. These pathways underlie diminished neuron growth that contributes to neurodevelopmental cortical circuit pathology. Thus, our experiments identify potential candidates for further development of mitochondria-based therapies and a molecular mechanistic framework for rational design of precisely targeted new drugs to correct molecular and cellular pathology associated with cortical neuron and circuits compromised in multiple neurodevelopmental disorders.