Molecular, cellular and physiological correlates of sustained attention in the locus coeruleus to anterior cingulate cortex circuit

项目总结 持续性注意力缺陷是神经精神障碍的一种突出的认知症状，如 精神分裂症、注意力缺陷多动障碍和严重抑郁障碍以及与年龄相关的 神经退行性疾病，包括阿尔茨海默病。在人类身上，连续性能测试(CPT) 通常用于评估患有这些神经精神疾病的患者的持续注意力。这个 前扣带回皮质(ACC)对于正常的CPT功能是必不可少的，患者群体经常表现出 CPT期间ACC功能异常。这些结果与行政协调会在以下方面的既定作用是一致的 注意引导行为，并指出了ACC在注意缺陷的病理生理学中的作用 复杂的脑部疾病。然而，支持ACC作用的细胞和分子机制 对持续关注的监管仍不清楚。这种知识的缺乏是很重要的，因为确定这些 机制对于开发针对注意力缺陷的靶向治疗至关重要。此应用程序调查 新的分子和细胞靶点的表达、ACC回路中的神经活动模式和 注意力表现。我们的初步数据表明，轨迹之间的投影特定路径 蓝斑(LC)和ACC调节CPT中注意力引导行为的不同方面。具体来说， 我们发现了编码载脂蛋白E(APOE)的基因，该基因与注意力和精神障碍有关 与以注意力缺陷为特征的障碍有关，作为潜在的分子参与者 持续关注LC-ACC电路。我们利用遗传和特定于电路的工具来剖析 在基于触摸屏的啮齿动物模拟过程中持续注意的分子、细胞和电路基础 人CPT(Rcpt)在小鼠体内的作用。具体地说，我们1)测试APOE基因表达与 LC-ACC回路中的生理功能和注意表现，以及2)识别细胞类型和回路- 啮齿动物和人类ACC中对持续注意力至关重要的特定分子靶点。要实现 这些目标，我们集成了各种分子和系统水平的方法，包括体内 电生理学、单细胞RNA测序和CRISPR-dCas9介导的表观基因组编辑 注意力引导行为的量化。对于测序研究，我们利用了 分子遗传学工具用于靶向和操纵LC-ACC回路中的细胞特异性群体 鼠标，并使用这些数据来从基因上识别死后人类数据中的电路特定细胞类型 脑组织。这种跨物种的分析支持了我们确定和优先治疗的长期目标 以注意力缺陷为特征的障碍的目标。这项拟议的研究意义重大，因为 将极大地促进我们对持续存在的电路和分子机制的理解 注意，以及为解剖学和基因定位的治疗靶向提供潜在途径 以注意力失调为特征的紊乱。

PROJECT SUMMARY Sustained attention deficits are a prominent cognitive symptom in neuropsychiatric disorders such as schizophrenia, attention-deficit hyperactivity disorder, and major depressive disorder as well as in age-related neurodegenerative disorders, including Alzheimer’s disease. In humans, continuous performance tests (CPTs) are commonly used to assess sustained attention in patients with these neuropsychiatric diseases. The anterior cingulate cortex (ACC) is essential for normal CPT performance, and patient populations often exhibit aberrant ACC function during the CPT. These results are in line with an established role for the ACC in attention-guided behavior, and point to a role for the ACC in the pathophysiology of attentional deficits in complex brain disorders. However, the cellular and molecular mechanisms that underlie the role of the ACC to regulate sustained attention remain unclear. This lack of knowledge is important because identifying these mechanisms is critical for developing targeted treatments for attention deficits. This application investigates links between expression of novel molecular and cellular targets, neural activity patterns in ACC circuits and attentional performance. Our preliminary data suggest that projection-specific pathways between the locus coeruleus (LC) and the ACC regulate distinct aspects of attention-guided behavior during the CPT. Specifically, we identified the gene encoding Apolipoprotein E (Apoe), which has been implicated in attention and disorders associated with disorders featuring deficits in attention, as a potential molecular player underlying regulation of sustained attention in the LC-ACC circuit. We leverage genetic and circuit-specific tools to dissect the molecular, cellular and circuit underpinnings of sustained attention during a touchscreen-based rodent analog of the human CPT (rCPT) in mice. Specifically, we 1) test causal relationships between Apoe gene expression, physiological function in the LC-ACC circuit and attentional performance, and 2) identify cell types and circuit- specific molecular targets in the rodent and human ACC that are critical for sustained attention. To achieve these aims we integrate a variety of molecular and systems level approaches including in vivo electrophysiology, single-cell RNA-sequencing, and CRISPR-dCas9 mediated epigenome editing coupled with quantification of attention-guided behavior. For the sequencing studies, we capitalize on the power of molecular genetic tools to target and manipulate cell-specific populations within the LC-ACC circuit in the mouse and use these data to genetically identify circuit-specific cell types in data from postmortem human brain tissue. This cross-species analysis supports our long-term goal of identifying and prioritizing therapeutic targets for disorders that feature attentional deficits. The proposed research is significant because the results will significantly advance our understanding of the circuit and molecular mechanisms underlying sustained attention, as well as provide potential avenues for anatomically and genetically-localized therapeutic targeting in disorders featuring dysregulation of attention.