Specification of sleep-wake control neurons in the basal forebrain

基底前脑睡眠-觉醒控制神经元的规范

• 批准号: 10454779
• 负责人: Ritchie Edward Brown
• 金额: --
• 依托单位: VA BOSTON HEALTH CARE SYSTEM
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-01 至 2024-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10454779
• 关键词: Ablation; Adult; Affect; Area; Arousal; Attention; Auditory; Autopsy; Behavior; Behavior Control; Bilateral; Brain; Cerebrum; Clinical Trials; Clozapine; Data; Dementia; Development; Developmental Biology; Diagnosis; Disease; Ectopic Expression; Electroencephalography; Electrophysiology (science); Embryo; Enterobacteria phage P1 Cre recombinase; Enzymes; FOXP2 gene; Future; Ganglia; Genes; Genetic; Genetic Techniques; Glutamates; Goals; Guide RNA; Homeobox; Human; Immunohistochemistry; Injections; Knock-in Mouse; Lateral; Lesion; Ligands; Link; Location; Maps; Medial; Methods; Military Personnel; Modeling; Motor Activity; Mouse Strains; Mus; Neurons; Neurotransmitters; Outcome; Oxides; Parvalbumins; Phenotype; Population; Predisposition; Preoptic Areas; Prosencephalon; Proteins; Reporter; Reporting; Research; Rewards; Risk; Role; Saline; Schizophrenia; Sleep; Sleep Disorders; Sleep Stages; Sleep Wake Cycle; Sleep disturbances; Sleeplessness; Specific qualifier value; Stains; Subgroup; Techniques; Testing; Time; Transcriptional Regulation; Transgenic Mice; Veterans; Viral Vector; Vulnerable Populations; Wakefulness; adeno-associated viral vector; alpha Toxin; basal forebrain; base; brain circuitry; cell type; cholinergic neuron; combat; density; design; experimental study; gamma-Aminobutyric Acid; genetic approach; innovation; interest; mind control; mouse Cre recombinase; neuropsychiatric disorder; neuropsychiatry; new therapeutic target; novel; novel strategies; outcome prediction; progenitor; programs; receptor; recombinase; response; risk variant; selective expression; sensory stimulus; sleep abnormalities; sleep regulation; sleep spindle; suicidal risk; targeted treatment; transcription factor; translational approach; translational study

Deployment and combat exposure substantially increase the risk for insomnia and other sleep disorders in military personnel. These findings match with the 6-fold increase in the diagnosis of sleep disorders in the VA in recent years. The most prevalent sleep disorder, insomnia, is associated with an increased risk for suicide. Furthermore, disrupted sleep and abnormal cortical activity are common in severe neuropsychiatric conditions affecting veterans such as dementia and schizophrenia. Thus, a better mechanistic understanding of the brain circuitry controlling sleep-wake cycles and cortical oscillations are urgently needed to develop novel treatments for veterans and other vulnerable populations. Accordingly, the broad objective of this research program in mice is to identify new therapeutic targets to correct abnormalities of sleep and cortical electrical activity. We will use an innovative approach in mice which characterizes subgroups of neurons based on their developmental origin and identifies the transcription factors which control their activity in adults, allowing targeted therapies which recalibrate their activity to restore normal sleep and cortical rhythms. We focus on the basal forebrain, a region involved in sleep-wake activity, attention and reward which degenerates in dementia, and on the largest group of neurons in this region which release the inhibitory neurotransmitter, gamma-amino-butyric acid (GABA). We target neurons which express three transcription factors linked to insomnia and other neuropsychiatric disorders common in veterans by genetic and postmortem studies. The roadmap to helping veterans is: We will identify new groups of neurons based on their developmental origin (Aim 1). We will use transgenic mice which allow manipulation of their activity to determine how they affect sleep-wake behavior and cortical oscillations (Aim 2). Finally, we will state-of-the-art genetic techniques to identify the transcription factors which control their activity in adults (Aim 3), allowing us to correct abnormal sleep-wake behavior and cortical electrical activity in neuropsychiatric disorders by designing viral vector based therapies which act on those transcription factors (long-term goal). Forebrain GABAergic neurons are generated in the subpallium, an area of the developing brain implicated in the risk for developing diverse neuropsychiatric disorders affecting veterans. Within the subpallium, different groups of GABAergic neurons are generated by progenitors in the caudal, medial and lateral ganglionic eminences and embryonic preoptic regions. In each region, different transcription factors specify unique groups of neurons. We will use genetically modified mice which express an enzyme, Cre recombinase (Cre), under the control of these transcription factors to identify them and manipulate their activity. Crossing these mouse strains with another mouse strain which expresses a red fluorescent protein in the presence of Cre, in combination with immunostaining will allow us to map their location and phenotype (Aim 1). In Aim 2 we will study their function by increasing their activity or ablating them using injections of viral vectors which express proteins in the presence of Cre. These experiments will reveal the role of these neurons in sleep-wake behavior and control of cortical electrical activity for the first time and set the stage for translational studies to alter their activity. Finally, in Aim 3, we will begin the first step towards a translational approach by using state-of-the art genetic techniques to modulate the activity of one of these transcription factors, Lhx6, whose expression is altered in a GABAergic cell-type implicated in schizophrenia. We will use a state-of-the-art gene editing technique called Clustered Regularly Interspersed Short Palindromic Repeats (CRISPR), which is currently being tested in clinical trials for various disorders. If successful, as suggested by our strong preliminary data, this project will allow novel translational approaches to study and correct disease-related abnormalities in veterans by manipulating the activity of basal forebrain neurons and genes which regulate sleep-wake behavior and cortical electrical activity.

部署和战斗暴露会大大增加失眠和其他睡眠障碍的风险 在军事人员中。这些发现与美国睡眠障碍诊断率增加了 6 倍相吻合。 近年来，VA。最普遍的睡眠障碍——失眠，与以下风险增加有关： 自杀。此外，睡眠中断和皮质活动异常在严重的神经精神疾病中很常见。 影响退伍军人的疾病，例如痴呆症和精神分裂症。因此，更好的机制理解 迫切需要开发控制睡眠-觉醒周期和皮质振荡的大脑电路 针对退伍军人和其他弱势群体的新疗法。因此，本次会议的总体目标 小鼠研究计划旨在确定新的治疗靶点，以纠正睡眠和皮质异常 电活动。我们将在小鼠身上使用一种创新方法，该方法基于神经元亚群的特征 确定其发育起源并确定控制其成人活动的转录因子， 允许有针对性的治疗重新调整其活动以恢复正常的睡眠和皮质节律。我们 关注基底前脑，该区域涉及睡眠-觉醒活动、注意力和奖励，但会退化 在痴呆症中，以及该区域释放抑制性神经递质的最大神经元群​​， γ-氨基丁酸（GABA）。我们的目标是表达三种转录因子的神经元 通过遗传和尸检研究，失眠和其他神经精神疾病在退伍军人中很常见。 帮助退伍军人的路线图是：我们将根据他们的神经元组来识别新的神经元组。 发展起源（目标 1）。我们将使用转基因小鼠，可以操纵它们的活动 确定它们如何影响睡眠-觉醒行为和皮质振荡（目标 2）。最后，我们将采用最先进的 遗传技术来识别控制成人活动的转录因子（目标 3），使我们能够 纠正神经精神疾病中异常的睡眠-觉醒行为和皮质电活动 设计基于病毒载体的疗法，作用于这些转录因子（长期目标）。 前脑 GABA 能神经元在大脑皮层下产生，这是大脑发育中的一个区域 涉及影响退伍军人的多种神经精神疾病的风险。内 在大脑皮层下，不同组的 GABA 能神经元由尾侧、内侧和内侧的祖细胞产生。 外侧神经节隆起和胚胎视前区。每个区域都有不同的转录因子 指定独特的神经元组。我们将使用表达酶 Cre 的转基因小鼠 重组酶（Cre），在这些转录因子的控制下识别它们并操纵它们 活动。将这些小鼠品系与另一种表达红色荧光蛋白的小鼠品系杂交 Cre 的存在与免疫染色相结合将使我们能够绘制它们的位置和表型 （目标 1）。在目标 2 中，我们将通过注射病毒来增加它们的活性或消除它们来研究它们的功能 在 Cre 存在的情况下表达蛋白质的载体。这些实验将揭示这些神经元的作用 首次在睡眠-觉醒行为和皮质电活动控制中发挥作用，并为 转化研究来改变他们的活动。最后，在目标 3 中，我们将开始迈向转化的第一步 通过使用最先进的遗传技术来调节这些转录之一的活性的方法 Lhx6 因子，其表达在与精神分裂症有关的 GABA 能细胞类型中发生改变。我们将使用一个 最先进的基因编辑技术，称为簇状规则散布短回文重复 （CRISPR），目前正在针对各种疾病进行临床试验。 如果成功，正如我们强有力的初步数据所表明的那样，该项目将实现新颖的转化 通过操纵基础活性来研究和纠正退伍军人疾病相关异常的方法 调节睡眠-觉醒行为和皮质电活动的前脑神经元和基因。