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.

项目总结/摘要 大约每400名儿童中就有1名出生时就有严重的听力损失，这使得先天性耳聋成为 影响幼儿的最常见疾病。大约50%的先天性耳聋是遗传性的， 起源目前，唯一可用的听力损失治疗方法是人工耳蜗植入或听力放大。 虽然这些治疗通常会改变生活，但它们在恢复正常听力方面的能力有限， 这导致了从童年开始的终生斗争。基因治疗方法 隐性听力损失带来了一个具有挑战性但令人兴奋的机会。将功能性基因病毒递送至 耳是具有挑战性的，特别是在小鼠中-目前的体内病毒递送方法只占一小部分， 感觉毛细胞是正常听力功能所必需的，并且只对较小的蛋白质容易起作用。此外，委员会认为， 可能需要病毒载体的多次应用以靶向治疗的最佳时机和持续时间。小鼠 缺乏肌动蛋白调节蛋白Eps 8的人类是耳聋的，而Eps 8基因敲除小鼠的静纤毛非常短 无法接触Corti器中的盖膜。一种新的转基因小鼠品系将 研究耳聋Eps 8 KO小鼠模型中出生后基因表达的潜力。使用 在PhyB系统中，将产生小鼠系，其中任何UAS控制的转基因的表达都可以被调控。 用红光激活或用远红光灭活。使用该系统，将实现UAS-Eps 8的体内表达 由红光或强力霉素诱导。系统地改变Eps 8表达的起始和持续时间， 则测试听功能和静纤毛伸长将有助于静纤毛的恢复潜力 在出生后小鼠体内的延长和听力恢复。此外，Eps 8在两种发展中的作用 以及静纤毛在生物体一生中的维持将在未来的研究中阐明。总的来说， 这一建议将为许多未来的项目铺平道路，这些项目将探索基因表达调控在人类基因组中的作用。 vivo将提供一个创新但实用的模型系统，用于探索和扩展关键时期， 听力恢复