GENERATION OF MODELS FOR GENETIC HEARING LOSS

描述（由申请人提供）：本提案的最终目标是 为最常见的听力损失类型创建猕猴模型。突变 连接蛋白26（GJB 2）基因中的突变是导致听力损失的最常见原因 （DFNB 1）在美国和欧洲人群中，约占所有 儿童听力损失目前尚无研究连接蛋白26的动物模型 因为敲除小鼠是胚胎致死的。预期 该项目的成果，猕猴连接蛋白26听力损失模型，将提供 一个重要的新的临床和基础研究的基础， 遗传性听力损失我们的假设是，基于 人类中平均约1/100的隐性基因，是产生模型的 通过筛选远系繁殖的灵长类动物来寻找人类隐性疾病将比 通过在小鼠中定向删除该基因。因为GJB 2基因的突变是 最常见的听力损失的遗传原因，我们选择了这个基因为我们的 初始屏幕。该序列在人和猕猴之间具有18个碱基的保守性 观察到的变化和4种氨基酸变化。我们将在一个 逐步实现以下具体目标。1)屏幕大 猕猴群体中GJB 2基因的突变。筛选将 优先和假定的突变功能将根据我们的评估 人类突变的经验。2)检测推定的GJB 2纯合子， 表型相关性一旦发现假定的病理性突变，交配或在 将安排在区域灵长类中心进行体外受精， 纯合子后代，并将提交研究表型的建议， 建立GJB 2猕猴群。3)生成猕猴模型， 听力损失基因灵长类动物之间的解剖和生理相似性 而人类也增加了灵长类动物模型的重要性。响应 猕猴的治疗将类似于人类，增加相关性， 临床研究。猕猴模型的基础听力损失研究将提供 深入了解导致人类听力损失的病理机制。 了解疾病过程将是设计和测试新的 治疗可以减少或预防人类遗传性听力损失。

DESCRIPTION (provided by applicant): The ultimate objective of this proposal is to create macaque models for the most prevalent types of hearing loss. Mutations in the gene for connexin 26 (GJB2) are the most common cause of hearing loss (DFNB1) in American and European populations, accounting for about 10% of all childhood hearing loss. An animal model is not available to study connexin 26 function in the ear because the mouse knockout is embryonic lethal. The expected outcome of this project, a macaque connexin 26 hearing loss model, will provide a foundation for important new clinical and basic studies of the most common type of inherited hearing loss. Our hypothesis, based on the carrier rate for the average recessive gene of about 1/100 in humans, is that generating a model for a recessive human disease by screening outbred primates will be easier than by targeted deletion of the gene in mice. Because mutations in GJB2 gene are the most common genetic cause of hearing loss, we have chosen this gene for our initial screen. The sequence is conserved between human and macaque with 18 base changes and 4 amino acid changes observed. We will test our hypothesis in a stepwise fashion by accomplishing the following Specific Aims. 1) Screen a large population of macaques for mutations in the GJB2 gene. Screening will be prioritized and putative mutation function will be evaluated based on our experience with human mutations. 2) Test the putative GJB2 homozygotes for phenotypic relevance. Once putative pathologic mutations are found, mating or in vitro fertilization at Regional Primate Centers will be arranged to produce homozygous offspring, and proposals will be submitted to study the phenotype and establish GJB2 macaque colonies. 3) Generate macaque models for additional hearing loss genes. Anatomical and physiological similarities between primates and humans have increased the importance of primate models. Responses of macaques to therapies will be similar to humans, increasing relevance of clinical studies. Basic hearing loss research in the macaque model will provide insights into pathologic mechanisms leading to hearing loss in humans. Understanding disease processes will be the basis for designing and testing new treatments that could reduce or prevent human inherited hearing loss.