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建立猕猴模型。突变 连接蛋白26(GJB2)的基因是导致听力损失的最常见原因 (DFNB1)在美国和欧洲人口中，约占所有人口的10% 儿童听力损失。尚无动物模型可用于研究连接蛋白26 在耳朵中起作用，因为老鼠的基因敲除是胚胎致命的。预期中的 这个项目的成果，猕猴连接蛋白26听力损失模型，将提供 为最常见疾病的重要新临床和基础研究奠定基础 遗传性听力损失类型。我们的假设是，基于 在人类中，平均约1/100的隐性基因是产生一个模型 对于隐性人类疾病，通过筛选异种灵长类动物将比 通过在小鼠身上定向删除该基因。因为GJB2基因的突变是 最常见的导致听力损失的基因，我们选择了这个基因 初始屏幕。该序列在人类和猕猴之间是保守的，具有18个碱基 观察到4种氨基酸的变化。我们将在一个 通过实现以下具体目标逐步实现时尚。1)大屏幕 GJB2基因突变的猕猴种群。放映将会是 优先排序和假定的突变功能将根据我们的 人类基因突变的经验。2)测试假定的GJB2纯合子 表型相关性。一旦发现假定的病理突变，交配或在 将安排在地区灵长类中心进行体外受精，以产生 纯合子后代，并将提交研究表型和 建立GJB2猕猴群体。3)为其他用户生成猕猴模型 听力损失基因。灵长类动物在解剖和生理上的相似性 人类增加了灵长类动物模型的重要性。的回应 猕猴的治疗将类似于人类，增加相关性 临床研究。猕猴模型中的基础听力损失研究将提供 对导致人类听力损失的病理机制的洞察。 了解疾病过程将是设计和测试新的 可以减少或防止人类遗传性听力损失的治疗方法。

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.