A Novel Model System for Restoring Hearing in vivo

一种恢复体内听力的新型模型系统

• 批准号: 10058225
• 负责人: Todd P Coleman
• 金额: $ 28.55万
• 依托单位: SALK INSTITUTE FOR BIOLOGICAL STUDIES
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10058225
• 关键词: Actins; Acute; Address; Adult; Affect; Age; Aging; Biological Models; Birth; Brain; Child; Childhood; Climacteric; Cochlear Hearing Loss; Cochlear Implants; Development; Disease; Doxycycline; Ear; Exhibits; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic; Hair Cells; Hearing; Hearing Aids; Hearing Tests; Human; Intestines; Knock-out; Knockout Mice; Labyrinth; Length; Light; Luciferases; Maintenance; Mechanics; Methods; Mus; Mutation; Neonatal; Newborn Infant; Organ of Corti; Organism; Photophobia; Physiological; Proteins; Repression; Resolution; Role; Sensory Hair; Signal Transduction; Structure; Switch Genes; System; Testing; Tetanus Helper Peptide; Therapeutic Studies; Tissue Extracts; Tissues; Transgenes; Transgenic Mice; Viral; Viral Vector; Wild Type Mouse; Work; base; congenital deafness; critical period; deaf; deafness; experimental study; gene induction; gene therapy; genetic regulatory protein; hearing impairment; hearing loss treatment; hearing restoration; hereditary hearing loss; high resolution imaging; imaging approach; improved; in vivo; innovation; insight; interest; mouse model; normal hearing; novel; phyA phytochrome; phyB phytochrome; postnatal; promoter; restoration; sound; spatiotemporal; tectorial membrane; therapy duration; tool; transduction efficiency; transgene expression

Project Summary/Abstract Approximately 1 out of 400 children are born with significant hearing loss, making congenital deafness one of the most common disorders affecting young children. Approximately 50% of congenital deafness is genetic in origin. Currently, the only available treatments for hearing loss are cochlear implants or hearing amplification. While these treatments are often life-changing, they are limited in their ability to restore hearing to normal, which results in lifelong struggles beginning acutely in childhood. Gene therapy approaches for treating recessive hearing loss presents a challenging but exciting opportunity. Viral delivery of functional genes to the ear is challenging, especially in mice – current in vivo viral delivery methods only transduce a fraction of the sensory hair cells necessary for proper hearing function, and only works easily for smaller proteins. Moreover, multiple applications of viral vectors may be required to target the optimal timing and duration for therapy. Mice and humans lacking the actin-regulatory protein Eps8 are deaf, and Eps8 KO mice have very short stereocilia that fail to contact the tectorial membrane in the organ of Corti. A novel transgenic mouse line will be generated to study the potential of postnatal gene expression in a deaf Eps8 KO mouse model. Using the PhyB system, a mouse line will be created wherein any UAS controlled transgene’s expression can be activated with red light or inactivated with far-red light. Using this system, UAS-Eps8 expression in vivo will be induced by either red light or doxycycline. Systematically varying the initiation and duration of Eps8 expression, then testing for hearing function and stereocilia elongation will facilitate the restoration potential of stereocilia elongation and hearing restoration in vivo in postnatal mice. Furthermore, the role of Eps8 in both development and maintenance of stereocilia over the lifetime of the organism will be elucidated in future studies. Overall, this proposal will pave the way for many future projects probing the effects of gene expression modulation in vivo and will provide an innovative but practical model system for probing and expanding the critical period for hearing restoration.

项目摘要/摘要 在400名儿童中，大约有1个天生有明显的听力损失，使先天性死亡成​​为 影响幼儿的最常见疾病。大约50％的先天性死亡是遗传的 起源。目前，听力损失的唯一可用治疗方法是人工耳蜗或听力放大。 尽管这些治疗经常改变生活，但它们的恢复听力正常的能力有限，但 这导致终生挣扎从童年开始急性。治疗基因治疗方法 隐性听力损失带来了一个挑战，但令人兴奋的机会。功能基因的病毒传递到 耳朵具有挑战性，尤其是在小鼠中 - 当前的体内病毒输送方法仅转导一小部分 感觉毛细胞需要适当的听力功能，仅适用于较小的蛋白质。而且， 可能需要使用病毒载体的多个应用来针对治疗的最佳时机和持续时间。老鼠 和缺乏肌动蛋白调节蛋白EPS8的人是聋的，EPS8 KO小鼠的立体胶质细胞非常短 那无法联系Corti器官中的技术膜。新颖的转基因鼠标系将是 生成以研究聋EPS8 KO小鼠模型中产后基因表达的潜力。使用 Phyb系统，将创建鼠标线，其中任何UAS控制转换的表达式可以是 用红光激活或用远红光灭活。使用此系统，在体内的UAS-EPS8表达将是 由红光或强力霉素诱导。系统地改变了EPS8表达的主动性和持续时间， 然后测试听力功能和立体核伸长率将促进立体丝菌的恢复潜力 产后小鼠体内的伸长和听力恢复。此外，EPS8在两种发展中的作用 在未来的研究中，将阐明生物体一生中立体柔性的维持。全面的， 该提案将为许多未来项目探索基因表达调制的影响 体内将提供一个创新但实用的模型系统，用于探测和扩展关键时期 听力恢复。