Hypocretin Gene Transfer and Narcolepsy

描述（由申请人提供）：嗜睡症现在被认为是一种神经退行性疾病，并且与CNS神经元死亡的其他疾病一样，有必要探索转移基因以恢复功能的新策略。在这里，我们建议开发一种基因转移的方法，将作为一种神经生物学工具，以了解网络基础嗜睡症，并最终扭转症状。我们已经创建了一个复制缺陷型HSV-1扩增子载体，将小鼠前下丘脑泌素原基因与报告基因一起转移到下丘脑泌素缺失小鼠中。我们非常强有力的初步数据显示，下丘脑分泌素缺失小鼠的外侧下丘脑中下丘脑分泌素丰富而强烈的表达，沿着发作性睡病症状的明显减轻。我们提出了一个综合系列的体外和体内的目标，将作为一个更全面的努力，利用基因转移的方法来扭转下丘脑泌素基因敲除小鼠嗜睡症的症状的基础。适当的控制实验，提出了加强的结论。在这里，我们专注于转移小鼠前下丘脑泌素原的基因，因为这种神经肽可以很容易地识别使用简单的免疫组织化学程序。然后，我们将转移受体的基因，这是一项更加困难的任务，因为没有好的抗体可以验证基因转移。我们的总体战略意图是转移犬发作性睡病中下丘脑泌素2受体的基因，从而用健康的受体基因取代突变的受体基因。发作性睡病现在被认为是一种神经退行性疾病，有必要探索治疗该疾病的新策略。该项目的意义在于，它将开发一种基因转移方法，该方法将作为一种神经生物学工具，用于了解嗜睡症的网络基础，并最终恢复某些功能。

DESCRIPTION (provided by applicant): Narcolepsy is now considered a neurodegenerative disorder and as with other diseases where CNS neurons die it is necessary to explore new strategies to transfer genes to restore function. Here we propose developing a gene transfer approach that will serve as a neurobiological tool to understand the networking underlying narcolepsy and also to ultimately reverse symptoms. We have created a replication-defective HSV-1 amplicon vector to transfer the gene for mouse preprohypocretin together with reporter genes into hypocretin null mice. Our very strong preliminary data shows abundant and robust expression of hypocretin in the lateral hypothalamus of hypocretin null mice along with unambiguous decline of narcoleptic symptoms. We are proposing an integrated series of in vitro and in vivo aims that will serve as a foundation for a more comprehensive effort to utilize the gene transfer approach to reverse the symptoms of narcolepsy in hypocretin null mice. Appropriate experiments with controls are proposed to strengthen the conclusions. Here, we are focusing on transferring the gene for mouse preprohypocretin because this neuropeptide can be easily identified using simple immunohistochemical procedures. We will then migrate to transferring the gene for the receptor, a much more difficult task since there is no good antibody that will allow for verification of the gene transfer. Our overall strategic intent is to transfer the gene for the hypocretin 2 receptor in canine narcolepsy, thereby replacing a mutated receptor gene with a healthy one. PUBLIC HEALTH RELEVANCE Narcolepsy is now considered a neurodegenerative disorder and it is necessary to explore new strategies to treat the disease. The significance of this project is that it will develop a gene transfer approach that will serve as a neurobiological tool to understand the networking underlying narcolepsy and also to ultimately restore some function.