Gene Transfer Into Selected Brain Neurons In Vivo

描述（由申请人提供）：病毒介导的基因转移的最新进展为神经科学研究提供了令人兴奋的新工具集。特别是在过去的10年里，许多研究已经使用病毒载体的显微注射将遗传物质插入到体内的大脑神经元中。然而，通常这些病毒操作并不是针对特定的细胞类型，而是针对注射部位内的所有细胞的病毒感染。此外，目前还没有使用这种技术明确调节神经元放电活动的研究。我们建议利用细胞型特异性病毒表达技术结合新的设计受体来克服这些限制。这项合作提案的目标是开发并验证一种方法，将设计受体插入体内大脑的食欲素神经元中，从而可以通过系统给药的化合物选择性地控制这一重要细胞群的冲动活动，否则在药理学上是惰性的。最近发现，食欲素在奖励和成瘾以及觉醒中都很重要。这项新技术的未来应用将包括选择性地调节食欲素神经元的活动，以更好地描述它们在成瘾中的作用。此外，选择性操纵特定细胞类型神经元的脉冲活动可以广泛应用于其他神经元，并将成为神经功能行为和电生理分析的重要新工具。拟议的研究与人类健康有关，因为它将开发新的工具，以更有选择性和特异性地操纵大脑神经元中的基因。这些新工具不仅对特定大脑神经元功能的基础研究具有重要意义，而且对新的治疗方法的开发也有重要意义，而且还将为大量神经系统疾病的基因治疗开辟新的临床方法。鉴于食欲素系统在成瘾和睡眠障碍中的作用，提出的具体方法将为控制食欲素神经元活动治疗成瘾或睡眠障碍（如嗜睡症）提供新的途径。

DESCRIPTION (provided by applicant): Recent advances in viral-mediated gene transfer have produced an exciting new toolset for neuroscience research. In particular, many studies in the last 10 years have used microinjection of viral vectors to insert genetic material into brain neurons in vivo. However, typically these viral manipulations have not been directed at a specific cell type, but rather viral infection has been targeted at all cells within the injection site. In addition, there have been almost no studies using such technology to explicitly modulate firing activity of neurons. We propose here to take advantage of cell-type specific viral expression technology coupled with new designer receptors to overcome these limitations. The goal of this collaborative proposal is to develop and validate a method to insert designer receptors into orexin neurons in brain in vivo, so that impulse activity in this important cell group can be selectively controlled by systemically administered compounds that are otherwise pharmacologically inert. Orexins have recently been found to be important in reward and addiction, as well as in arousal. Future applications of this new technology would include regulation of activity in orexin neurons selectively to better delineate their role in addiction. In addition, selective manipulation of impulse activity in cell- type specific neurons can be applied widely to other neurons, and will be a valuable new tool in behavioral and electrophysiological analyses of neural function. PUBLIC HEALTH RELEVANCE The proposed research is relevant to human health because it will develop new tools for more selective and specific manipulation of genes in brain neurons. These new tools not only will be important for basic research into the functions of specific brain neurons with consequences for new therapeutic development, but will also lead the way for new clinical approaches in gene therapy for a large number of disorders of the nervous system. Given the roles of the orexin system in addiction and in sleep disorders, the specific method proposed will lead to new ways to control orexin neuron activity for treating addiction or sleep disorders such as narcolepsy.