Astroglial Orexin in Sleep Disorders

睡眠障碍中的星形胶质细胞食欲素

• 批准号: 8585663
• 负责人: Priyattam J. Shiromani
• 金额: $ 22.43万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8585663
• 关键词: Affect; Animal Disease Models; Animal Model; Area; Astrocytes; Axon; Behavior; Brain; Brain region; Calcium; Cataplexy; Cations; Chemicals; Complex; Corpus striatum structure; Data; Development; Disease; Distal; Gene Delivery; Gene Transfer; Genes; Genetic; Genetic Engineering; Glial Fibrillary Acidic Protein; Hour; Image; In Vitro; Knock-out; Leg; Length; Ligands; Light; Link; MJD1 protein; Measures; Methods; Modeling; Motor; Movement; Mus; Narcolepsy; Nerve Degeneration; Neurobiology; Neuroglia; Neurons; Obstructive Sleep Apnea; Optics; Peptides; Pharmacologic Substance; Pharmacology; Pontine structure; Publications; Research; Role; Site; Sleep; Sleep Disorders; Specificity; Symptoms; Testing; Transgenic Mice; behavior change; cost; cost effective; hypocretin; improved; mammilloinfundibular nucleus structure; mouse model; novel strategies; optogenetics; promoter; public health relevance; receptor; research study; synaptic function; tool

DESCRIPTION (provided by applicant): We have demonstrated the first successful use of neuronal-orexin gene transfer to ameliorate symptoms of narcolepsy in two established animal models of the disease. The effects were site specific and depended on the connectivity of the surrogate neurons. Since neuronal orexin can be secreted at multiple distal sites, some of which may regulate narcoleptic behavior whereas others do not, we propose a localized gene delivery method by expressing orexin in astroglia. Astroglial-orexin will re-establish the ligand-receptor link in a localized area that already contains the orexin receptors. Astroglial-orexin gene deliver will serve as a cost-effective neurobiological tool to understand the networking underlying sleep. In aim 1 the gene for orexin will be inserted into astroglia (GFAP promoter driven) in three specific brain regions, two of which are implicated in network sleep models (TMN, pons) and the third, striatum, will serve as control. Our preliminary results in the orexin-ataxin-3 mice model indicate that in the TMN glial-orexin (rAAV-GFAP-orexin) completely rescues waking but not cataplexy. In the pons it produces a 78% reduction in cataplexy but not waking. Thus, there is site- specificity of the glial-orexin. In aim 2 astroglial-orexin will be driven by optogenetic stimulation to determine whether such stimulation further stimulates waking or blocks cataplexy, especially in ineffective sites in aim 1. Experiments with appropriate controls, including orexin receptor antagonist are proposed to strengthen the conclusions. In-vitro calcium imaging study will determine functionality of the genetically inserted ChR2 receptors. To the best of our knowledge this is the first use of astroglial-orexin in sleep disorders. Gene transfer is extremely cost-efficient, it is used clinically and it will demonstrate that genetic pharmacology can be used to elucidate the network in sleep disorders.

描述（由申请人提供）：我们已经在两种已建立的动物模型中证明了首次成功使用神经元食欲素基因转移来改善发作性睡病的症状。效果是位点特异性的，并且取决于替代神经元的连接性。由于神经元食欲素可以在多个远端部位分泌，其中一些可以调节发作性睡病行为，而另一些则不能，因此我们提出了一种通过在星形胶质细胞中表达食欲素的局部基因递送方法。星形胶质细胞食欲素将在已经含有食欲素受体的局部区域重新建立配体-受体连接。星形胶质细胞食欲素基因传递将作为一种经济有效的神经生物学工具来了解睡眠背后的网络。在目标 1 中，食欲素基因将被插入到三个特定大脑区域的星形胶质细胞（GFAP 启动子驱动）中，其中两个与网络睡眠模型（TMN、脑桥）有关，第三个纹状体将作为对照。我们在 orexin-ataxin-3 小鼠模型中的初步结果表明，在 TMN 中，神经胶质食欲素 (rAAV-GFAP-orexin) 完全可以挽救清醒，但不能挽救猝倒。在脑桥中，它可以减少 78% 的猝倒症状，但无法使人清醒。因此，神经胶质食欲素具有位点特异性。在目标 2 中，星形胶质细胞食欲素将由光遗传学刺激驱动，以确定这种刺激是否进一步刺激清醒或阻止猝倒，特别是在目标 1 中无效的部位。建议使用适当的对照（包括食欲素受体拮抗剂）进行实验，以加强结论。体外钙成像研究将确定基因插入的 ChR2 受体的功能。据我们所知，这是星形胶质细胞食欲素首次用于治疗睡眠障碍。基因转移极其 成本效益高，用于临床，将证明遗传药理学可以使用 阐明睡眠障碍中的网络。