Hypocretin Gene Transfer and Narcolepsy

描述(申请人提供)：发作性睡病现在被认为是一种神经退行性疾病，与其他中枢神经系统神经元死亡的疾病一样，有必要探索新的策略来转移基因以恢复功能。在这里，我们建议开发一种基因转移方法，作为一种神经生物学工具来理解发作性睡病的潜在网络，并最终扭转症状。我们已经建立了一个复制缺陷的HSV-1扩增载体，将小鼠前降钙素基因和报告基因一起转移到下丘脑素缺失的小鼠中。我们非常强大的初步数据显示，在下丘脑下丘脑外侧，下丘脑中有大量和强劲的下丘脑表达，伴随着发作性睡病症状的明确下降。我们提出了一系列综合的体外和体内目标，将作为更全面的努力的基础，以利用基因转移方法来逆转下丘脑泌素缺失小鼠的发作性睡病症状。为了加强这一结论，提出了适当的对照实验。在这里，我们把重点放在转移小鼠前原分泌素的基因上，因为这种神经肽可以通过简单的免疫组织化学程序很容易地识别出来。然后我们将转移到受体的基因转移，这是一个困难得多的任务，因为没有好的抗体来验证基因转移。我们的总体战略意图是在犬类发作性睡病中转移下丘脑分泌素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。与公共卫生相关的发作性睡病现在被认为是一种神经退行性疾病，有必要探索治疗这种疾病的新策略。这个项目的意义在于，它将开发一种基因转移方法，作为一种神经生物学工具来理解发作性睡病背后的网络，并最终恢复一些功能。

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.