Targeting Mitochondrial Function to Develop Novel Therapies for Neurodevelopmental Disorders

摘要 我们将确定潜在的新的药理学疗法，以改善一类线粒体功能 大脑皮层神经元的损伤被认为是多种神经发育障碍的损害。我们有 显示在LgDel小鼠的第2/3层投射神经元（PN）中线粒体代谢被破坏 22q11.2缺失综合征（一种综合征性神经发育障碍）模型。我们还证明 这种破坏显然解释了关联皮层连接的数量差异， 与LgDel小鼠的认知行为缺陷相关。最后，我们发现自由基清除剂 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种线粒体靶向药物的能力， 化合物，以及已知的靶向药物，以调节LgDel层2/3 PN线粒体 代谢及其对2/3层PN稳态的影响。在具体目标1中，我们将评估化合物 使用Agilent Seahorse代谢分析仪进行的经验证的均匀层2/3 PN体外试验中的活性 来测量线粒体功能的关键方面。恢复LgDel线粒体的候选化合物 在不破坏WT线粒体功能的情况下， 减少异常的线粒体活性氧（ROS）水平，恢复树突和轴突 在LgDel层2/3 PN中生长。为了提供机械精度的额外解释性解决方案， 这些化合物，在特定目标2中，我们将进行LgDel与WT的平行转录组比较 体外2/3层PN，以鉴定由于线粒体DNA而改变其转录调节的途径。 神经发育病理学靶向的发育中的第2/3层PN的功能障碍和生长减少。 我们将进一步将该数据与用NAC处理的LgDel层2/3 PN的转录组谱进行对比， 代谢和生长恢复活性，我们以前已经证明。因此，本发明中鉴定的化合物 将验证筛选的特异性、靶向细胞活性，并将其置于差异调节的背景下。 对线粒体功能改变敏感的转录途径。这些途径导致了 导致神经发育皮层回路病理的神经元生长。因此，我们的实验确定 潜在的候选人，为进一步发展的药物为基础的治疗和分子机制 合理设计精确靶向新药以纠正分子和细胞病理学的框架 与多种神经发育障碍中受损的皮质神经元和回路相关。

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.