Identification of Genes Causing Syndromic and Nonsyndromic Hearing Impairment

导致综合征性和非综合征性听力损伤的基因的鉴定

• 批准号: 8565496
• 负责人: Thomas Friedman
• 金额: $ 343.73万
• 依托单位: NATIONAL INSTITUTE ON DEAFNESS AND OTHER COMMUNICATION DISORDERS
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8565496
• 关键词: 15q21; 16p; 19p; Adult; Affect; Antibodies; Auditory system; Boston; Calcium-Binding Proteins; Cell Line; Cellular biology; ChIP-seq; Chromosomes; Clinical; Collaborations; Development; Family; Female; Gene Structure; Genes; Genetic; Genetic Markers; Genetic Variation; Goals; Gonadal Dysgenesis; Hearing; Human; Human Genetics; Inherited; Laboratories; Labyrinth; Link; Lung; Maps; Methodology; Molecular; Molecular Biology; Mus; Mutate; Mutation; Nature; Pathway interactions; Perrault syndrome; Phenotype; Proteins; Publishing; Pulmonary alveolar structure; Regulation; Reporting; Surveys; Technology; Tissues; Type II Epithelial Receptor Cell; Universities; Usher Syndrome; Work; cell type; deafness; gene function; genome wide association study; hearing impairment; interest; lung development; medical schools; mouse model; next generation; novel; recessive genetic trait; research study; tool; transcription factor

The goal of the Section on Human Genetics is to identify and study the function of mutated genes for human hereditary deafness. This work begins with the ascertainment of large families in which deafness appears to be inherited either as a dominant or a recessive trait. We then search for linkage of the deafness to genetic markers for the known (already reported) syndromic, DFNA (dominant) and DFNB (recessive) loci. If linkage to the known deafness loci can be excluded, we initiate a genome-wide screen to search for novel deafness loci followed by work to identify the causative gene. During the past year we ascertained several large families segregating deafness, mapped a novel deafness locus, and identified a novel gene for nonsyndromic deafness. Sections of the following projects were completed in the past year and have been published or are in press and likely to be published in 2012. 1. We recently mapped a novel nonsyndromic deafness locus (DFNB81) to chromosome 19p, which is distinct from the closely linked DFNB72 locus (Rehman et al., 2011 EJHG). Mutations of GIPC3 are responsible for DFNB72 deafness. In collaboration with Drs. Suzanne Leal and William Newman, we are working to identify the mutated gene underlying DFNB81 deafness using next-generation sequencing technology. DFNB81 deafness was initially presumed to be nonsyndromic. However, further clinical chactareization of the affected subjects indicates Perrault syndrome characterized by hearing loss and female gonadal dysgenesis. 2. We mapped a novel locus for nonsyndromic deafness to chromosome 16p. The locus is designated DFNB86 (Ali et al., 2011). We are working to identify the mutated gene underlying DFNB86 deafness gene using advanced sequencing methodologies. 3. Grhl2-TMinsC/+ is a mouse model of DFNA28 human progressive hearing loss, which we reported ten years ago (Peters et al., 2002). A goal of our present study is to understand the function of the GRHL2 transcription factor in the auditory system. To that end, we have performed ChIP-Seq experiments using our own and commercially available antibodies to GRHL2, in a variety of tissues and cell types. In collaboration with Drs. Maria Ramirez and Saaket Varma at Boston University Medical School, and as a way to validate this technology in our laboratory, we extended our ChIP and expression analyses to include developing lung buds and the MLE15 cell line derived from adult mouse lung. GRHL2 is required for normal lung development. Varma et al. 2012, (Morell and Friedman are co-authors) showed that GRHL2 and NKX2-1 transactive each other and form a regulatory loop defining the critical transition from a type II cell type (cuboidal) to type I (squamous) in the lung alveolus. We are making a more comphrehensive survey of GRHL2 regulatory targets in the lung as compared to the auditory system using ChIP-Seq and our AB5500 sequencer. 5. Nonsyndromic deafness DFNB48 and a novel type 1 Usher syndrome were previously genetically mapped by us to an overlapping interval on chromosome 15q21-q23 (Ahmed et al., 2009). We recently demonstrated that mutations associated with DFNB48 and USH1J are allelic and that the causative gene encodes a calcium binding protein (Riazuddin et al., 2012, Nature Genetics, in press).

人类遗传学部分的目标是识别和研究人类遗传性耳聋突变基因的功能。这项工作首先确定大家庭中耳聋似乎是作为显性或隐性特征遗传的。然后，我们寻找耳聋与已知（已报道）综合征、DFNA（显性）和 DFNB（隐性）基因座的遗传标记的联系。 如果可以排除与已知耳聋基因座的连锁，我们将启动全基因组筛选来寻找新的耳聋基因座，然后确定致病基因。在过去的一年里，我们确定了几个分离耳聋的大家族，绘制了一个新的耳聋基因座，并鉴定了一个新的非综合征性耳聋基因。 以下项目的部分内容已于去年完成，并已出版或正在出版，并可能于 2012 年出版。 1. 我们最近将一个新的非综合征性耳聋基因座 (DFNB81) 定位到染色体 19p，该基因座不同于紧密连锁的 DFNB72 基因座 (Rehman et al., 2011 EJHG)。 GIPC3 突变导致 DFNB72 耳聋。与博士合作。 Suzanne Leal 和 William Newman，我们正在利用下一代测序技术来鉴定导致 DFNB81 耳聋的突变基因。 DFNB81 耳聋最初被认为是非综合征性的。然而，受影响受试者的进一步临床特征表明佩罗综合征，其特征是听力损失和女性性腺发育不全。 2. 我们将非综合征性耳聋的一个新基因座定位到染色体 16p。该基因座被指定为 DFNB86（Ali 等人，2011）。我们正在努力使用先进的测序方法来识别 DFNB86 耳聋基因的突变基因。 3. Grhl2-TMinsC/+ 是 DFNA28 人类进行性听力损失的小鼠模型，我们十年前报道过该模型（Peters 等，2002）。我们目前研究的目标是了解 GRHL2 转录因子在听觉系统中的功能。为此，我们使用我们自己的和市售的 GRHL2 抗体在多种组织和细胞类型中进行了 ChIP-Seq 实验。与博士合作。波士顿大学医学院的 Maria Ramirez 和 Saaket Varma，作为在我们实验室验证这项技术的一种方法，我们扩展了 ChIP 和表达分析，以包括发育中的肺芽和源自成年小鼠肺的 MLE15 细胞系。 GRHL2 是正常肺部发育所必需的。瓦尔玛等人。 2012 年，（Morell 和 Friedman 为共同作者）表明 GRHL2 和 NKX2-1 相互交互并形成一个调节环路，定义肺泡中从 II 型细胞类型（立方形）到 I 型（鳞状）细胞的关键转变。与听觉系统相比，我们正在使用 ChIP-Seq 和我们的 AB5500 测序仪对肺部 GRHL2 调控靶点进行更全面的调查。 5. 非综合征性耳聋 DFNB48 和一种新型 1 型 Usher 综合征先前被我们通过基因映射到染色体 15q21-q23 上的重叠区间（Ahmed 等，2009）。我们最近证明，与 DFNB48 和 USH1J 相关的突变是等位基因，并且致病基因编码钙结合蛋白（Riazuddin 等人，2012，Nature Genetics，出版中）。