In Vivo Genetic Manipulation of Neuronal Excitability

神经元兴奋性的体内遗传操作

• 批准号: 6711042
• 负责人: David Clifford Yeomans
• 金额: $ 16.92万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-03-05 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6711042
• 关键词: Alphaherpesvirinae; C fiber; Herpesviridae; SDS polyacrylamide gel electrophoresis; action potentials; behavior test; chronic pain; confocal scanning microscopy; flow cytometry; fluorescence microscopy; gene expression; genetic regulation; immunocytochemistry; laboratory mouse; neural information processing; neural transmission; neurons; sodium channel; transfection /expression vector; voltage /patch clamp; western blottings

DESCRIPTION (provided by applicant): Despite many years of research, the mainstays of clinical treatment of pain remain highly problematic. High dose opiate analgesics have numerous side effect, induce tolerance and dependence and have significant abuse potential. New knowledge and technologies, however, may allow for entirely new approaches to the management of chronic pain. It has recently been established that there are 12 or more different isoforms of neuronal sodium channels, which are critical to these neurons' ability to convey information. One isoform in particular, Nav1.7, appears to be selectively present in neurons that carry pain information from the periphery to the central nervous system. Furthermore, levels of these channels, as well as their location on these neurons, change dramatically during chronic pain states, implying that they are important in establishing and/or maintaining these pain states. Thus, an examination of Nav1.7 channel function may be significant in designing new approaches to the treatment of chronic pain. In order to establish the importance of this sodium channel in pain, virally directed gene transfer will be used to decrease or eliminate expression of the gene for Nav1.7 selectively in those neurons that carry pain information. Herpes viral vectors have been previously used to insert novel transgenes into these neurons, and to decrease synthesis of endogenous proteins, but this approach has never been applied to sodium channels. Thus, a herpes virus encoding an antisense sequence for the Nav1.7 channel gene will be used to knock-down production of this channel, so that immunochemical, behavioral, and electrophysiological experiments can be conducted in order to examine the contribution of Nav1.7 in pain biology. In this way, it should become clear whether or not this channel represents a potential target for the development of new pharmaceutical tools. Another implication of this work, however, is that this approach may in itself provide a means of therapy, and it may be possible to use herpes vectors to inhibit production of endogenous Nav1.7 channels in chronic pain patients, providing a long-term genetic therapy for their pain.

描述（由申请人提供）： 尽管多年的研究，疼痛的临床治疗的支柱仍然存在很大的问题。大剂量阿片类镇痛药副作用大，易产生耐受性和依赖性，有被滥用的危险。然而，新的知识和技术可能会为慢性疼痛的管理提供全新的方法。 最近已经确定有12种或更多种不同的神经元钠通道亚型，这些亚型对这些神经元传递信息的能力至关重要。特别是一种亚型，Nav1.7，似乎选择性地存在于将疼痛信息从外周传递到中枢神经系统的神经元中。此外，这些通道的水平，以及它们在这些神经元上的位置，在慢性疼痛状态期间显著变化，这意味着它们在建立和/或维持这些疼痛状态中是重要的。因此，Nav1.7通道功能的检查在设计治疗慢性疼痛的新方法中可能是重要的。 为了确定这种钠通道在疼痛中的重要性，将使用病毒定向基因转移来选择性地减少或消除携带疼痛信息的那些神经元中Nav1.7基因的表达。疱疹病毒载体先前已被用于将新的转基因插入这些神经元中，并减少内源性蛋白质的合成，但这种方法从未应用于钠通道。因此，编码Nav1.7通道基因的反义序列的疱疹病毒将被用于敲低该通道的产生，从而可以进行免疫化学、行为和电生理学实验以检查Nav1.7在疼痛生物学中的贡献。通过这种方式，应该清楚这种渠道是否代表了开发新药物工具的潜在目标。然而，这项工作的另一个意义是，这种方法本身可能提供一种治疗手段，并且有可能使用疱疹载体来抑制慢性疼痛患者内源性Nav1.7通道的产生，为他们的疼痛提供长期的基因治疗。