Gene Therapy for Hearing and Balance Disorders

听力和平衡障碍的基因疗法

• 批准号: 10222650
• 负责人: DAVID P COREY
• 金额: $ 42.3万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10222650
• 关键词: Animals; Antibody titer measurement; Auditory; Behavior; Biological Assay; Blindness; Brain Stem; Capsid; Cell physiology; Cells; Clarin-1; Cochlea; Complement; Data; Ear; Electron Microscopy; Electrophysiology (science); Gene Delivery; Gene Transduction Agent; Gene Transfer; Genes; Genotype; Goals; Guidelines; Hair Cells; Hearing; Hearing problem; Hereditary Disease; Human; Immune response; Immunity; In Vitro; Injections; Labyrinth; Lead; Measures; Mediating; Membrane; Mus; Mutation; Operative Surgical Procedures; Pathogenicity; Patients; Perilymph; Phenotype; Preclinical Testing; Primates; Sampling; Serotyping; Serum; T cell response; Testing; Toxic effect; Toxicity Tests; United States Food and Drug Administration; Usher Syndrome Type 3A; Variant; Viral; Viral Vector; Work; adeno-associated viral vector; deafness; design; efficacy testing; equilibration disorder; gene therapy; genetic deafness; hearing impairment; hearing restoration; immunogenicity; improved; in vivo; induced pluripotent stem cell; meetings; mouse model; mutant; mutant mouse model; neutralizing antibody; nonhuman primate; novel; otoacoustic emission; restoration; round window; transduction efficiency; variant of unknown significance; vector

Project Summary This project will develop a gene therapy strategy to treat hearing loss associated with Usher Syndrome type 3A (USH3A) in human. This will involve optimizing viral vectors in mouse models of USH3A, testing the best vectors for efficacy and cochlear toxicity in non-human primates and in human hair cells, and testing sequence variants in the human USH3A gene (CLRN1) for pathogenicity. At the completion of the project, we will be able to request a pre-IND meeting with the Food and Drug Administration to determine specific requirements for preclinical testing. Aim 1 is to test different AAV vectors for gene delivery to cochlear hair cells in mouse. Three new AAV- related vectors have shown promise in transducing cochlear hair cells, and we will test them all to identify the best. We will then test AAV-mediated restoration of hair cell function in the Clrn1-/- and N48K mouse models. Finally, we will test toxicity to hearing of AAV-mediated Clrn1 expression. Aim 2 is to test the efficacy of novel AAV vectors in human hair cells in vitro, and to test, in non-human primates, the efficiency, toxicity and immunogenicity of these vectors administered by round window membrane injection. We will test the two best vectors from Aim 1, expressing GFP to assess the degree of viral transduction of hair cells. We will also describe toxicity and systemic distribution of GFP- and CLRN1- encoding AAV vectors. An immune response might interfere with subsequent AAV injections or could lead to toxicity in the inner ear, so we will determine whether AAV injection into the ear induces neutralizing antibody or T-cell response. In Aim 3 we will characterize human pre-existing immunity against applicable serotypes in perilymph, using perilymph collected during surgery for other auditory disorders. We will also determine whether there is a correlation between antibody titers in perilymph and serum. Many variants have been identified in the CLRN1 gene but not all are known to be pathogenic. To determine which variants in CLRN1 in USH3A patients are pathogenic, we will set up a complementation assay using the Clrn1-/- mouse, use AAV to express Clrn1 with specific mutations equivalent to human variants of unknown significance, and assess degree of functional rescue with electrophysiology and electron microscopy.

项目摘要 该项目将开发一种基因治疗策略来治疗与Usher综合征3A型相关的听力损失 （USH3A）。这将涉及在USH 3A小鼠模型中优化病毒载体，测试最佳的 在非人灵长类动物和人毛细胞中的效力和耳蜗毒性的载体，以及测试顺序 在人类USH 3A基因（CLRN 1）的致病性变异。在项目完成后，我们将 能够要求与食品药品监督管理局召开IND前会议，以确定具体要求 用于临床前测试。 目的1：研究不同的腺相关病毒载体对小鼠耳蜗毛细胞的基因转染作用。三个新的AAV- 相关的载体在转导耳蜗毛细胞方面显示出了希望，我们将对它们进行测试，以确定 最好然后，我们将在Clrn 1-/-和N48 K小鼠模型中测试AAV介导的毛细胞功能恢复。 最后，我们将测试AAV介导的Clrn 1表达对听力的毒性。 目的2是测试新型AAV载体在体外人毛细胞中的功效，并测试在非人毛细胞中的功效。 灵长类动物，通过圆窗施用这些载体的效率、毒性和免疫原性 膜注射我们将测试来自Aim 1的两个最佳载体，表达GFP以评估病毒感染的程度。 毛细胞的转导。我们还将描述GFP-和CLRN 1-的毒性和全身分布。 编码AAV载体。免疫反应可能会干扰随后的AAV注射或可能导致 因此，我们将确定AAV注射到耳中是否诱导中和 抗体或T细胞反应。 在目标3中，我们将使用以下方法来表征针对外淋巴液中适用血清型的人预先存在的免疫力： 手术期间收集的外淋巴液用于治疗其他听觉疾病。我们还将确定是否有一个 外淋巴和血清中抗体滴度之间的相关性。许多变种已被确定在 CLRN 1基因，但不是所有已知的致病性。为了确定USH 3A中CLRN 1的哪些变体 患者是致病性的，我们将使用Clrn 1-/-小鼠建立互补测定，使用AAV表达 clrn 1的特定突变相当于人类变异的未知意义，并评估程度， 用电生理学和电子显微镜进行功能拯救。