Autosomal Dominant Non-Syndromic Hearing Loss - Its Genetic Diagnosis and Treatment

项目摘要 听力损失影响着世界上15-26%的人口。在老年人中，它是最常见的 神经系统残疾，影响约50%的老年人。大多数遗传性迟发性耳聋是 常染色体显性遗传和非综合征型。在这个新的阶段，我们将利用我们的经验和专业知识， 人类遗传学和基因治疗，以改善遗传性耳聋的耳聋，通过利用单一的力量， 细胞RNA测序，并确定靶向等位基因抑制是否可以预防或潜在逆转 通过使用RNA干扰（RNAi）在人类耳聋的成熟动物模型中观察听力损失。 为实现这些目标，我们提出以下具体目标： ·具体目标1.使用单细胞分离技术，我们建议： o SA1A。为了完善我们对Corti器官中单个细胞特异性转录的认识， 通过鉴定和表征新外显子和亚型进行耳聋的综合基因检测 与非综合征型和综合征型听力损失有关的基因。 o SA1B。为了描述单个基因突变对个体转录组的更广泛影响， 人ADNSHL的两种鼠模型-Kncq 4 +/-小鼠，其是人ADNSHL的模型， DFNA 2相关的听力损失，以及Tmc 1Bth/+小鼠（贝多芬（Bth）小鼠），这是一个模型， 人DFNA 36相关的听力损失。 ·具体目标2。使用RNAi，我们建议： 〇 SA2A。确定靶向等位基因抑制是否可以预防或潜在逆转听力损失 在成熟的动物中。 〇 SA2B。制定一个广泛适用的治疗策略，用于由单一原因引起的所有类型的耳聋。 使用新的RNAi构建体抑制内源等位基因和伴随基因 在一些实施方案中，所述基因被工程化以抵抗RNAi介导的沉默的外源野生型等位基因取代。 o SA2C。为了确定针对AAV的系统性中和抗体（NAb）是否影响转导， AAV介导的耳蜗基因治疗的效率。 这些目标的成功实现将促进我们对听觉生物学的理解， 耳聋，加强对听力损失患者的临床护理，并可能为临床 使用RNAi作为某些类型听力损失的治疗选择的试验。

Project Summary Hearing loss affects 15-26% of the world's population. Amongst the elderly, it ranks as the most common neurological disability, impacting ~50% of octogenarians. The majority of inherited late-onset deafness is autosomal dominant and non-syndromic. In this new porposal, we will build on our experience and expertise in human genetics and gene therapy to improve genetic deafness for deafness by leveraging the power of single- cell RNA sequencing and to establish whether targeted allele suppression can prevent or potentially reverse hearing loss in mature animal models of human deafness by using RNA interference (RNAi). To achieve these goals, we propose the following specific aims: • Specific Aim 1. Using single-cell isolation techniques, we propose: o SA1A. To refine our knowledge of individual cell-specific transcription in the organ of Corti and improve comprehensive genetic testing for deafness by identifying and characterizing novel exons and isoforms of genes implicated in both non-syndromic and syndromic forms of hearing loss. o SA1B. To characterize the broader impact of single gene mutations on the transcriptome of individual cells in two murine models of human ADNSHL – the Kncq4+/- mouse, which is a model of human DFNA2-related hearing loss, and the Tmc1Bth/+ mouse (the Beethoven (Bth) mouse), which is a model of human DFNA36-related hearing loss. • Specific Aim 2. Using RNAi, we propose: o SA2A. To establish whether targeted allele suppression can prevent or potentially reverse hearing loss in mature animals. o SA2B. To develop a broadly applicable treatment strategy for all types of deafness caused by a single gene using a novel RNAi construct that suppresses both endogenous alleles with concomitant gene replacement by an exogenous wild-type allele engineered to resist RNAi-mediated silencing. o SA2C. To determine whether systemic neutralizing antibodies (NAbs) to AAV impact the transduction efficiency of AAV-mediated cochlear gene therapy. The successful completion of these aims will advance our understanding of the biology of hearing and deafness, enhance the clinical care of persons with hearing loss, and potentially lay the foundation for clinical trials using RNAi as a treatment option for some types of hearing loss.