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：
10460137
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/10460137
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.

项目概要该项目将开发一种基因治疗策略来治疗与 3A 型亚瑟综合症相关的听力损失（USH3A）在人类中。这将涉及优化 USH3A 小鼠模型中的病毒载体，测试最佳的病毒载体用于非人类灵长类动物和人类毛细胞中的功效和耳蜗毒性的载体以及测试序列人类 USH3A 基因 (CLRN1) 的致病性变异。项目完成后，我们将能够请求与食品和药物管理局举行 IND 前会议以确定具体要求用于临床前测试。目标 1 是测试不同的 AAV 载体将基因传递至小鼠耳蜗毛细胞。三个新的 AAV- 相关载体在转导耳蜗毛细胞方面已显示出前景，我们将对它们进行全部测试以识别最好的。然后，我们将在 Clrn1-/- 和 N48K 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 Clrn1 表达对听力的毒性。目标 2 是在体外测试新型 AAV 载体在人类毛细胞中的功效，并在非人类毛细胞中进行测试灵长类动物，通过圆窗施用这些载体的效率、毒性和免疫原性膜注射。我们将测试目标 1 中表达 GFP 的两个最佳载体，以评估病毒感染程度毛细胞的转导。我们还将描述 GFP- 和 CLRN1- 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 注射或可能导致内耳毒性，因此我们将确定AAV注射入耳中是否会引起中和抗体或 T 细胞反应。在目标 3 中，我们将使用以下方法描述人类针对外淋巴中适用血清型的预先存在的免疫力：在其他听觉障碍手术期间收集的外淋巴。我们还将确定是否存在外淋巴液和血清中抗体滴度的相关性。已发现许多变种 CLRN1 基因但并非所有基因都已知具有致病性。确定 USH3A 中 CLRN1 的哪些变异患者是致病性的，我们将使用 Clrn1-/- 小鼠建立互补实验，使用 AAV 表达 Clrn1 具有相当于未知意义的人类变异的特定突变，并评估其程度通过电生理学和电子显微镜进行功能性救援。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

AAV-S: A versatile capsid variant for transduction of mouse and primate inner ear.

AAV-S：一种多功能衣壳变体，用于转导小鼠和灵长类动物内耳。

DOI：
发表时间：
2021-06-11
期刊：
影响因子：
0
作者：
Ivanchenko, Maryna V;Hanlon, Killian S;Hathaway, Daniel M;Klein, Alex J;Peters, Cole W;Li, Yaqiao;Tamvakologos, Panos I;Nammour, Josette;Maguire, Casey A;Corey, David P
通讯作者：
Corey, David P