Hypocretin and its receptors Gene Transfer for Narcolepsy

描述（由申请人提供）：我有一个非凡的旅程。我在中国北京以北一千英里的一个小镇上长大。我在中国的教育体系中一路向上，获得了医学博士学位。和博士在中国最好的医科大学：北京医科大学。2002年，我加入了阿尔弗雷德·盖勒博士在哈佛医学院/退伍军人医学中心的实验室。经过4年的分子生物学和基因治疗培训，我于2006年加入Priyattam Shiromani博士的实验室，开始了一个新的研究领域：睡眠障碍的基因治疗。我们于2008年发表了嗜睡症基因转移研究领域的第一篇“主要证据”论文。我被提升为讲师，这是哈佛医学院的一个教师级别的职位，也是在那一年，我获得了永久居民身份。2011年，我们的实验室搬迁到南卡罗来纳州医科大学，我被晋升为助理教授（终身教职）。我打算在美国发展我的学术载体。我精通分子生物学，并对神经科学有广泛的了解。然而，为了达到我的职业目标，我需要在睡眠神经生物学方面进行额外的培训，因为这对我来说是一个新的领域。 这项提案的长期目标是开发一种治疗人类嗜睡症和其他睡眠障碍的基因疗法。短期目标是确定导致发作性睡病症状的特定HCRT靶区域及其表型。这项建议将提供睡眠神经生物学，特别是嗜睡症的培训。培训包括在既定科学家的监督下进行基本的实验室工作，课程工作，科学会议上的演讲，以及访问既定研究人员以了解其他睡眠障碍和新方法，如光遗传学。这项培训将使我成为睡眠神经科学的独立科学家，并使我在获得独立的NIH研究资助（R 01）方面具有竞争力。 嗜睡症是一种神经退行性睡眠障碍，影响近1：2000的美国人。十年前，神经肽下丘脑泌素（HCRT），又称食欲素（orexin），被认为与发作性睡病有关，HCRT缺乏被认为是人类发作性睡病的主要原因。但目前还不清楚HCRT的关键靶点在大脑的哪个部位。这些信息是必要的，以确定潜在的治疗，以扭转发作性睡病症状。目前对发作性睡病的治疗主要依靠药物治疗，这些药物可以治疗发作性睡病的部分症状，但不是全部症状，并且大多数药物都有严重的副作用。我们的团队是嗜睡症基因治疗领域的先驱。我们已经确定，HCRT基因转移在外侧下丘脑（LH）或其他相关的大脑区域，如Zona Incerta可以纠正发作性睡病的症状HCRT/共济失调蛋白-3转基因小鼠，一个可靠的小鼠模型发作性睡病。基于这项工作的成功，我们提出了这个项目，我们将构建载体，将HCRT或其受体的基因转移到参与睡眠-觉醒调节的特定大脑区域，或用光遗传学方法刺激/抑制这些神经元。我们的战略意图是使用基因转移和光遗传学工具来逆转嗜睡症的症状，从而确定调节睡眠的大脑区域。我们将研究这种基因转移策略是否也可以纠正老年（2岁）小鼠的发作性睡眠症状。我是唯一一个在睡眠研究中对老年小鼠进行基因转移研究的人。我相信识别出拯救发作性睡眠行为的替代神经元的表型是很重要的。这种靶向治疗正在被用于其他临床疾病，我的愿景是， 也可用于治疗发作性睡病和其他睡眠和觉醒障碍的特定症状。该项目的结果将为发现更好的替代方案奠定坚实的基础 人类嗜睡症的治疗方法 公共卫生相关性：我们将开发一种基因递送方法，将下丘脑泌素或其受体的特异性基因转移到下丘脑泌素/共济失调蛋白-3转基因或HCRT受体缺失小鼠中。这种“遗传药理学”方法与最新的光遗传学技术相结合，代表了一种神经生物学工具，可以了解嗜睡症背后的神经网络，嗜睡症是一种神经退行性睡眠障碍，与含有神经肽下丘脑泌素（也称为食欲素）的神经元丢失有关。该项目的结果将为发现更好的人类嗜睡症替代疗法奠定坚实的基础。这一建议将作为一个坚实的桥梁，成为一个独立的研究者在睡眠神经生物学和其他神经系统疾病。

DESCRIPTION (provided by applicant): I have had a remarkable journey. I grew up in a small town about a thousand miles north of Beijing, China. I worked my way up through the Chinese system, obtaining both M.D. and Ph.D. in the best medical university in China: Beijing Medical University. I then joined Dr. Alfred Geller's laboratory at the Harvard Medical School/VA Medical Center in 2002. After 4 years training in molecular biology and gene therapy, I joined Dr. Priyattam Shiromani's laboratory in 2006 to start a new research field: gene therapy for sleep disorders. We published the first "proof of principal" paper in narcolepsy gene transfer research area in 2008. I was promoted to instructor, a faculty level position at Harvard Medical School and also that year I received my permanent resident status. In 2011, our laboratory relocated to the Medical University of South Carolina and I was promoted to assistant professor (tenure track). I am intent on developing my academic carrier in the US. I am skilled in molecular biology and have a broad overall knowledge of neuroscience. However, for me to reach my career objectives I require additional training in sleep neurobiology, as this is a new field for m. The long-term goal of this proposal is to develop a gene therapy treatment for human narcolepsy and other sleep disorders. The short term goal is to determine the specific HCRT target areas responsible for narcoleptic symptoms, and their phenotypes. This proposal will provide training in sleep neurobiology, especially narcolepsy. Training includes basic lab work under the supervision of established scientists, course work, presentations at scientific meetings, and visiting established investigators to learn about other sleep disorders and new methods such as optogenetics. This training will enable me to become an independent scientist in sleep neuroscience and allow me to be competitive in securing an independent NIH research grant (R01). Narcolepsy is a neurodegenerative sleep disorder affecting almost 1:2000 Americans. Ten years ago the neuropeptide hypocretin (HCRT), also known as orexin, was linked to narcolepsy, and deficiency of HCRT was found to be the main reason of human narcolepsy. But it is still not known where in the brain are the key targets for HCRT. This information is necessary to identify potential therapies to reverse the narcoleptic symptoms. Current treatments of narcolepsy mainly depend on pharmacotherapy which can treat some but not all of the symptoms of narcolepsy and most of these medicines have severe side effects. Our group is pioneering in the field of Narcolepsy gene therapy. We have established that HCRT gene transfer in lateral hypothalamus (LH) or other related brain areas such as Zona Incerta can correct narcoleptic symptoms in HCRT/ataxin-3 transgenic mice, a reliable mouse model of narcolepsy. Drawing on the success of this work we are proposing this project where we will construct vectors to transfer the genes for the HCRT or its receptor into specific brain areas involved in sleep-wake regulation, or stimulate/inhibit these neurons with the optogenetic method. Our strategic intent is to use gene transfer and optogenetic tools to reverse the symptoms of narcolepsy and thereby identify the brain area that regulates sleep. We will examine if such gene transfer strategies can also correct narcoleptic symptoms in aged (2y) mice. I am the only one in sleep research conducting gene transfer studies in old mice. I believe it important to identify the phenotype of the surrogate neuron that rescues the narcoleptic behavior. Such targeted therapy is being used in other clinical disorders, and my vision is that it can also be used to treat specific symptoms of narcolepsy and other disorders of sleep and wakefulness. Results of this project will serve as a strong base for discovering better alternative treatments for human narcolepsy. PUBLIC HEALTH RELEVANCE: We will develop a gene delivery method to transfer specific genes for the hypocretin or its receptor into hypocretin/ataxin-3 transgenic or HCRT receptor-null mice. Such a "genetic pharmacology" method, combined with the newest optogenetic technique, represents a neurobiological tool to understand the neural networking underlying narcolepsy, a neurodegenerative sleep disorder linked to the loss of neurons containing the neuropeptide hypocretin, also named orexin. Results of this project will serve as a strong base for discovering better alternative treatments for human narcolepsy. This proposal will work as a solid bridge towards becoming an independent investigator in sleep neurobiology and other neurological diseases.