GENE TRANSFER AND CELL DEATH IN THE COCHLEA

耳蜗中的基因转移和细胞死亡

• 批准号: 6564019
• 负责人: YEHOASH RAPHAEL
• 金额: $ 15.76万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-12-01 至 2002-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6564019
• 关键词: Adenoviridae; BCL2 gene /protein; basal ganglia; brain stem; cell death; cochlea; electrophysiology; gene expression; guinea pigs; transfection; transfection /expression vector; virus genetics; virus infection mechanism

This proposal will investigate gene transfer and cell death in the cochlea. Gene transfer has recently become an important method for tissue engineering for experimental and clinical purposes. The experimental field of inner ear biology and the otology clinic may benefit greatly from gene transfer technology. Viral vectors have become the vehicle of choice for in vivo genetic interventions in the nervous system. The cochlea may be among the organs that can benefit from genetic manipulation via viral vectors. We have demonstrated efficient infection of guinea pig spiral ganglion cells in vitro and in vivo with the Ad.RSVntlacZ adenoviral vector. Transgene expression has been detected as long as eight weeks after the inoculation. In this application we propose to continue our successful preliminary experiments to optimize variable parameters of adenoviral mediated gene transfer into cochlear cells. Specifically, we will (a) determine the optimal inoculation parameters needed for efficient infection of spiral ganglion cells, (b) characterize the immune response to the viral infection, (c) determine the duration of viral gene expression, (d) characterize the spread of infection to adjacent tissues, and (e) determine the functionality of infected spiral ganglion cells using ABR measurements. We also propose to determine the effects of gene transfer on spiral ganglion cell survival. Specifically, we will determine if it is possible to (a) rescue spiral ganglion cells by overexpressing bcl-2 and (b) induce cell death in spiral ganglion cells by viral-mediated overexpression of bcl-xs. The data we have been generating, along with the experiments we now propose will solidify the foundation of viral mediated gene transfer in the cochlea. This technology will facilitate experimental elimination of specific cell types in the cochlea, and create a powerful tool for physiological studies in this organ. As such, it will be of benefit to the other projects described in our Program Project. Moreover, gene transfer will enable to study function of specific genes in the ear. It will also help the important clinical goal of preserving spiral ganglion cells, for reducing progressive trauma and enhancing benefits from cochlear implants.

该建议将研究基因转移和细胞死亡 耳蜗。基因转移最近已成为组织的重要方法 用于实验和临床目的的工程。 实验 内耳生物学领域和耳鼻喉科诊所可能会从中受益匪浅 基因转移技术。病毒向量已成为首选的工具 用于神经系统中的体内遗传干预措施。耳蜗可能 成为可以通过病毒受益的器官之一 向量。 我们已经证明了有效感染豚鼠螺旋神经节 带有AD.RSVNTLACZ腺病毒载体的体外和体内细胞。 转基因表达已被检测到后长达八周 接种。 在此应用程序中，我们建议继续我们的成功 初步实验以优化腺病毒的可变参数 介导的基因转移到耳蜗细胞中。 具体来说，我们将（a） 确定有效的最佳接种参数 螺旋神经节细胞的感染，（b）表征免疫反应 为了病毒感染，（c）确定病毒基因的持续时间 表达，（d）表征感染传播到相邻组织的传播， （e）确定感染的螺旋神经节细胞的功能 使用ABR测量。我们还建议确定基因的影响 转移螺旋神经节细胞存活。具体来说，我们将确定 如果有可能（a）通过过表达来拯救螺旋神经节细胞 Bcl-2和（b）通过病毒介导的螺旋神经节细胞诱导细胞死亡 BCl-XS的过表达。 我们一直在生成的数据以及现在的实验 建议将巩固病毒介导的基因转移的基础 耳蜗。这项技术将有助于实验消除 耳蜗中的特定细胞类型，并为 该器官的生理研究。因此，这将有益于 我们计划项目中描述的其他项目。此外，基因转移 将能够研究耳朵中特定基因的功能。 它也会 帮助保留螺旋神经节细胞的重要临床目标 减少进行性创伤并增强人工耳蜗的益处。