MOLECULAR GENETICS OF AND GENE THERAPY FOR DEAFNESS

耳聋的分子遗传学和基因治疗

• 批准号: 2443547
• 负责人: Anil K Lalwani
• 金额: $ 11.85万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-07-01 至 2001-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2443547
• 关键词: adeno associated virus group; beta galactosidase; clinical research; cochlea; congenital deafness; disease /disorder model; disease /disorder proneness /risk; drug delivery systems; family genetics; gene therapy; genetic markers; guinea pigs; human subject; immunocytochemistry; in situ hybridization; linkage mapping; neural degeneration; neurogenetics; neurotrophic factors; nucleic acid sequence; transfection; transfection /expression vector

Hereditary hearing impairment affects between 0.05 to 0.1% of the total US population. However, the precise cause of inherited auditory impairment at the molecular level is poorly understood due to the genetic heterogeneity of deafness. A strategy that overcomes the problem with genetic heterogeneity involves carrying out linkage analysis on families with nonsyndromic hereditary deafness. A three generation family with Schiebe or cochleosaccular degeneration, the most common cause of profound, congenital hearing impairment accounting for approximately 70% of cases of hereditary deafness, that is suitable for linkage studies has been identified. The immediate goal of the proposed research project is to map the location of gene responsible for Scheibe degeneration in this family using commercially available genomic markers. Once linkage to a region of the human genome is established, suitable candidate genes that reside in the linked region will be identified. These candidate genes will then be searched for the pathological mutation. The genetic study of this family and the subsequent identification of the gene which, when mutated, results in Scheibe degeneration should greatly aid in the understanding of the development and homeostatic mechanisms of the inner ear. The purpose of the second component of this proposal is to assess the utility of adeno-associated virus (AAV, a non-pathogenic human parvovirus) in introducing genetic material into the neuroepithelia of the inner ear. While gene transfer has been successfully performed in a large variety of post-mitotic cells such as myotubes, hepatocytes, endothelial cells, airway epithelial cells, and a variety of neuronal cells, the inner ear has not been studied as a target for gene therapy. The AAV containing the bacterial beta-galactosidase (beta-gal) sequence, a marker gene whose product is readily detectable, will be infused selectively into the cochlea with the aid of an osmotic minipump. The infused cochleas will be studied at various interval-transduction to assess the specificity and stability of AAV infection. Experimentally, the ability to introduce genes into the inner ear will aid in the understanding the function of cochlear proteins and control of inner ear specific genes. Therapeutically, the prompt delivery of neurotrophic factors could reduce the tissue damage and preserve hearing following injury. Positional cloning studies in several large families with hereditary, non-syndromic hearing impairment represent potential candidate genes for gene therapy. These studies will aid in designing therapeutic strategies to alleviate auditory dysfunction as well as contributing towards molecular genetic analysis of hearing.

遗传性听力障碍影响总数的 0.05% 至 0.1% 美国人口。 然而，遗传性听觉的确切原因 由于遗传因素，我们对分子水平的损伤知之甚少。 耳聋的异质性。 克服该问题的策略 遗传异质性涉及对家系进行连锁​​分析 患有非综合征性遗传性耳聋。 一个三代家庭 Schiebe 或耳蜗囊变性，最常见的原因 严重的先天性听力障碍约占 70% 适合进行关联研究的遗传性耳聋病例 已被识别。 拟议研究项目的近期目标是 绘制负责 Scheibe 变性的基因的位置 家庭使用商业上可用的基因组标记。 一旦链接到 人类基因组区域确定后，合适的候选基因 位于链接区域的将被识别。 这些候选基因 然后将搜索病理突变。 遗传学研究 这个家族的基因以及随后的基因鉴定，当 突变，Scheibe 变性的结果应该极大地有助于 了解内部的发展和稳态机制 耳朵。 本提案第二部分的目的是评估 腺相关病毒（AAV，一种非致病性人类病毒）的效用 细小病毒）将遗传物质引入神经上皮细胞 内耳。 虽然基因转移已成功进行 多种有丝分裂后细胞，如肌管、肝细胞、 内皮细胞、气道上皮细胞和多种神经元 细胞，尚未将内耳作为基因治疗的靶点进行研究。 含有细菌β-半乳糖苷酶（β-gal）序列的AAV， 其产物易于检测的标记基因将被注入 借助微型渗透泵选择性地进入耳蜗。 这 输注的耳蜗将以不同的时间间隔进行研究 评估 AAV 感染的特异性和稳定性。 实验上， 将基因引入内耳的能力将有助于 了解耳蜗蛋白的功能和内耳的控制 特定基因。治疗上，及时给予神经营养剂 因素可以减少组织损伤并保持听力以下 受伤。 几个大家族的定位克隆研究 遗传性、非综合征性听力障碍代表潜在的 基因治疗的候选基因。 这些研究将有助于设计 缓解听觉功能障碍的治疗策略 有助于听力的分子遗传学分析。