Gene Editing for DFNA9 Deafness

DFNA9 耳聋的基因编辑

• 批准号: 10389933
• 负责人: Hila Romi
• 金额: $ 7.96万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-12-06 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10389933
• 关键词: Gene; Editing; DFNA9; Deafness

PROJECT SUMMARY/ABSTRACT Significant advances in gene discovery in the field of hearing and deafness have raised a great interest in developing new therapeutic approaches for genetic deafness. Hearing loss (HL), is the most common sensory deficit worldwide, affecting the quality of life of people of all ages. Despite the progress in the understanding of genetic etiologies and the molecular mechanisms involved in deafness, no effective therapies in restoring the natural hearing are as yet available. Given the massive burden posed by hearing problems and the emerging landscape of gene editing technology, it is now essential to address the unmet medical need and to pursue new strategies for treatment of inherited hearing loss. Recent reports of preclinical studies for developing targeted therapeutic approaches for inherited deafness have opened promising avenues for precision medicine in hearing loss. Recent reports have yielded encouraging results in preclinical trials of virally mediated gene therapy for inherited deafness. One disorder of interest is DFNA9, an autosomal dominant nonsyndromic late-onset HL with variable vestibular dysfunction, caused by mutations in COCH, encoding cochlin, the most abundantly detected protein in the inner ear. This disease model is similar to and representative of the majority of the genetic HL disorders with a dominant mode of inheritance with an underlying deleterious gain-of-function/dominant-negative mechanism of the mutant protein. The goal of this project is to develop a gene therapy strategy to treat hearing loss associated with DFNA9, along with training of a research fellow in the fields of human genetics of deafness. To achieve these goals, we plan to utilize state-of-the-art CRISPR-Cas9 gene editing technology to develop an allele-specific targeting approach, and to test its efficacy in halting or mitigating the development or progression of hearing loss in a knock-in (KI) mouse model of DFNA9 associated with the dominant COCH p.A449T mutation. Preliminary in vitro studies support the specificity of the developed CRISPR-Cas9 reagents and strategies. Furthermore, additional very recent phenotypic characterization of our novel Coch A449T KI mouse model at eight to nine months is consistent with late-onset hearing loss. We plan to perform more in-depth phenotyping of the Coch A449T KI mouse model, and to explore the mechanism of pathogenicity. We further plan to evaluate the efficiency and specificity of in vivo CRISPR-Cas9 target gene disruption in prevention or mitigation of progressive hearing loss. Carrying out the project aims will provide substantial progress in development of the essential steps required for translation of gene therapy into clinical application. This project will establish a knowledgeable research fellow in the field of hereditary deafness and gene therapy, who plans to lead research in precision medicine for hearing loss.

项目总结/摘要 听力和耳聋领域基因发现的重大进展引起了人们对以下方面的极大兴趣： 为遗传性耳聋开发新的治疗方法。听力损失（HL）是最常见的感觉 这是世界范围内的赤字，影响到所有年龄段的人的生活质量。尽管在理解方面取得了进展， 遗传病因学和耳聋的分子机制，没有有效的治疗方法来恢复 自然听觉还没有出现。考虑到听力问题和新出现的 随着基因编辑技术的发展，现在必须解决未满足的医疗需求，并寻求新的 治疗遗传性听力损失的方法最近的临床前研究报告， 遗传性耳聋的治疗方法为听力精确医学开辟了有希望的途径 损失最近的报道在病毒介导的基因治疗的临床前试验中取得了令人鼓舞的结果， 遗传性耳聋一种感兴趣的疾病是DFNA 9，一种常染色体显性非综合征性晚发型HL， 可变前庭功能障碍，由编码cochlin的COCH突变引起， 内耳中的蛋白质这种疾病模型与大多数遗传性HL相似并具有代表性。 具有潜在有害功能获得性/显性阴性的显性遗传模式的疾病 突变蛋白的机制。该项目的目标是开发一种基因治疗策略来治疗听力 与DFNA 9相关的损失，沿着培训人类耳聋遗传学领域的研究员。 为了实现这些目标，我们计划利用最先进的CRISPR-Cas9基因编辑技术来开发一种 等位基因特异性靶向方法，并测试其在停止或减轻发展或进展中的功效。 在与显性COCH p.A449T突变相关的DFNA 9基因敲入（KI）小鼠模型中， 初步的体外研究支持所开发的CRISPR-Cas9试剂和策略的特异性。 此外，我们的新型Coch A449 T KI小鼠模型的额外的非常新的表型表征， 八到九个月是晚发性听力损失我们计划进行更深入的表型分析 Coch A449 T KI小鼠模型的致病性，并探讨其致病机制。我们计划进一步评估 体内CRISPR-Cas9靶向基因破坏在预防或减轻 渐进性听力损失项目目标的实现将在发展 将基因治疗转化为临床应用所需的基本步骤。该项目将建立一个 在遗传性耳聋和基因治疗领域知识渊博的研究员，他计划领导研究 听力损失的精准治疗