Molecular Genetics on Non Syndromic Hearing Loss (NSHL)

摘要：听力损失是影响2800多万美国人的最常见的感觉障碍。临床上 在出生时，至少每500名婴儿中就有1名出现显著的HL。近70%出生时表达的HL有遗传病因。 至少80%的非综合征性耳聋是常染色体隐性遗传(ARNSHL)。这些基因的鉴定已经 极大地改善了聋人和重听家庭的临床诊断和管理。然而，有一个 迫切需要继续鉴定新的人类HL基因和确定已知NSHL基因的致病变异 完成了一个基因组和表型数据库。这一点至关重要，因为我们和其他人已经提供了直接的证据 HL的进一步遗传异质性与一些基因/突变尚未确定：致病变异在 不到40%的病例和最近的研究表明，超过1000个基因与耳聋有关，这表明 在人类和小鼠中，许多导致耳聋的基因仍有待鉴定。鉴定所有HL基因 致病突变对于我们理解正常听力和疾病的生物学来说是必不可少的。 病因学/过程，为参与咨询和诊断的家庭提供直接好处，并在分子水平上 水平，允许开发新的基因特异性甚至突变特异性疗法来治疗HL。后者通过使用 基因组编辑可以有更广泛的用途，而不仅仅是在该基因突变的家庭中。重要的是，如图所示 在我们的初步研究中，我们已经从一个大型国际队列中收集了dna样本和表型数据。 (迈阿密耳遗传学资料库)NSHL家庭，建立了良好的迈阿密耳科诊所和管道 对变异的遗传和功能分析，成功地排除了大量多基因家族中所有已知的HL基因 确定了许多潜在的新候选基因，并为这些人建立了几个具有耳聋表型的动物模型 使用CRISPR/Cas9系统和传统的定向突变方法检测耳聋基因。在这项提案中，我们将 在我们以前的成就和初步数据的基础上，建议完成以下具体目标：1) 确定已知NSHL基因中的致病变异，以扩展组合基因组和表型耳聋数据库； 2)利用靶向序列捕获/全外显子(WES)/基因组(WGS)分析鉴定新的ARNSHL基因 使用定制的本地管道平台；3)通过体外和体外实验确定耳聋基因的功能后果 使用创新方法的活体模型。我们将表演一场最大、最完整的 迄今对ARNSHL的临床/基因组/功能研究。这项创新的研究不仅将增加我们对 听力和耳聋的生物学，但将高度翻译，导致改善的病因诊断 NSHL和患者护理以及通过在NSHL中生成的动物模型为基因/变体特异性治疗铺平道路 求婚。

Abstract: Hearing loss (HL) is the most common sensory disorder affecting more than 28 million Americans. Clinically significant HL is present in at least 1 per 500 infants at birth. Nearly 70% of HL expressed at birth has a genetic etiology. At least 80% of nonsyndromic deafness is autosomal recessive (ARNSHL). The identification of these genes has dramatically improved the clinical diagnosis and management of deaf and hard-of-hearing families. However, there is a pressing need to continue identifying new human HL genes and to determine causative variants in known NSHL genes for completing a genomic and phenotypic database. This is critical as we and others have provided direct evidence of further genetic heterogeneity of HL with a number of genes/mutations yet to be identified: causative variants are found in less than 4 0 % of cases and recent studies suggest that over a thousand genes are involved in deafness suggesting there are many deafness-causative genes still remaining to be identified in both human and mouse. Identifying all of the HL genes and causative mutation is imperative to our understanding of the biology of normal hearing and the disease etiology/processes, provides immediate benefit to the families involved for counseling and diagnosis, and, at the molecular level, allows for the development of novel gene-specific and even mutation-specific therapies to treat HL. The latter by using genome editing that can have much wider use than just in the families with mutations in that gene. Importantly, as shown in our preliminary studies, we have already collected DNA samples and phenotype data from a large international cohort (Miami Otogenetic Repository) of families with NSHL, well established the Miami Otogenetic Clinic and pipelines for the genetic and functional analysis of variants, excluded all known HL genes in a large cohort of multiplex families, successfully identified many potential new candidate genes, and generated several animal models with deafness phenotype for these human deafness genes using the CRISPR/Cas9 system and traditional targeted mutagenesis approach. In this proposal, we will build on our previous accomplishments and preliminary data by proposing to complete the following specific aims: 1) to determine causative variants in known NSHL genes for expanding a combined genomic and phenotypic deafness database; 2) to identify novel ARNSHL genes using targeted sequence capture/ whole exome (WES)/genome (WGS) analysis using a customized local pipeline platform; 3) to determine functional consequences of deafness genes with in vitro and in vivo models using innovative approaches. We will perform one of the largest and most integrated clinical/genomic/functional studies on ARNSHL to date. This innovative study will not only increase our understanding of the biology of hearing and deafness, but will be highly translational leading to improvements in the etiological diagnosis of NSHL and patient care as well as pave the wave for gene/variant specific treatments with animal models generated in the proposal.