Gene Therapy for Hearing and Balance Disorders

听力和平衡障碍的基因疗法

基本信息

批准号：
9978805
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
项目类别：
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/9978805
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

项目摘要

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.

项目摘要该项目将制定一种基因疗法策略，以治疗与3A型Usher综合征相关的听力损失（USH3A）在人类中。这将涉及在USH3A的鼠标模型中优化病毒向量，测试最佳非人类灵长类动物和人毛细胞中有效性和人工耳蜗毒性的载体，并测试序列人类USH3A基因（CLRN1）的变体用于致病性。项目完成时，我们将能够要求与食品药品监督管理局进行预先开会以确定特定要求用于临床前测试。目的1是测试不同的AAV矢量，以将基因递送到小鼠中的耳蜗细胞。三个新的AAV- 相关矢量已经在转导耳蜗细胞方面显示出希望，我们将测试它们以识别最好的。然后，我们将测试CLRN1 - / - 和N48K小鼠模型中毛细胞功能的AAV介导的恢复。最后，我们将测试听到AAV介导的CLRN1表达的毒性。 AIM 2是测试新型AAV载体在体外人毛细胞中的功效，并在非人类中测试这些向量的灵长类动物，效率，毒性和免疫原性。膜注射。我们将测试AIM 1的两个最佳向量，表达GFP以评估病毒的程度毛细胞的转导。我们还将描述GFP和CLRN1-的毒性和系统分布编码AAV向量。免疫反应可能会干扰随后的AAV注射，或者可能导致内耳的毒性，因此我们将确定AAV注射到耳朵中是否诱导中和抗体或T细胞反应。在AIM 3中，我们将表征人类预先存在针对Perilymph中适用血清型的免疫力，并使用在其他听觉疾病的手术期间收集的围绕膜。我们还将确定是否有 perilymph和血清中的抗体滴度之间的相关性。在 CLRN1基因，但并非全部都是致病性的。确定USH3A中CLRN1中的哪些变体患者是病原性的，我们将使用CLRN1 - / - 小鼠建立一个补充测定法，使用AAV表达 CLRN1具有与未知意义的人类变异的特异性突变，并评估程度用电生理学和电子显微镜进行功能救援。