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。特别是，在过去的10年中，许多研究已经使用病毒载体的显微注射将遗传物质插入体内脑神经元中。然而，通常这些病毒操作并不针对特定的细胞类型，而是病毒感染靶向注射部位内的所有细胞。此外，几乎没有研究使用这种技术来明确地调节神经元的放电活动。我们建议利用细胞类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和验证一种方法，将设计受体插入体内大脑中的食欲素神经元，以便可以通过全身给药的化合物选择性地控制这一重要细胞群的冲动活动，否则这些化合物是惰性的。食欲素最近被发现在奖励和成瘾以及唤醒中很重要。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性，以更好地描述它们在成瘾中的作用。此外，对细胞类型特异性神经元脉冲活动的选择性操纵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一种有价值的新工具。公共卫生相关性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脑神经元功能的基础研究具有重要意义，并对新的治疗方法的发展产生影响，而且还将为大量神经系统疾病的基因治疗新的临床方法开辟道路。鉴于食欲素系统在成瘾和睡眠障碍中的作用，提出的具体方法将导致控制食欲素神经元活性的新方法，用于治疗成瘾或睡眠障碍，如嗜睡症。

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.