Fetal Pharmacotherapy for Congenital Deafness

先天性耳聋的胎儿药物治疗

• 批准号: 9275471
• 负责人: JOHN Vincent BRIGANDE
• 金额: $ 32.73万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-18 至 2019-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9275471
• 关键词: 5' Splice Site; Address; Adenoviruses; Affect; Age; Antisense Oligonucleotides; Area; Auditory; Auditory Threshold; Base Pairing; Behavior assessment; Bilateral; Biological Models; Birth; Blindness; Child; Clinic; Clinical; Cochlea; Cochlear Implants; Collaborations; DNA Sequence Alteration; Defect; Dose; Embryology; Epithelial; Epithelium; Equilibrium; Excitatory Amino Acids; Fetal Therapies; Frequencies; Functional disorder; GTP-Binding Protein alpha Subunits, Gs; Gene Expression; Gene Transduction Agent; Gene Transfer; Genes; Genetic; Glutamates; Goals; Hair Cells; Hearing; Hearing Aids; Hearing Impaired Persons; Human; Image-Guided Surgery; Individual; Inherited; Inner Hair Cells; Intervention; Knockout Mice; Labyrinth; Live Birth; Mediating; Messenger RNA; Microinjections; Missense Mutation; Modeling; Molecular; Molecular Analysis; Molecular Virology; Morphology; Mus; Mutant Strains Mice; Mutation; National Institute on Deafness and Other Communication Disorders; Neurons; Neurotransmitters; Nucleotides; Pathogenicity; Pathway interactions; Pharmacotherapy; Physiological; Proteins; Protons; RNA Splicing; RNA, Messenger, Splicing; Research Priority; Retinitis Pigmentosa; SLC17A8 gene; Satellite Viruses; School-Age Population; Sensory; Sensory Hair; Severities; Site; Structure; Synaptic Transmission; Synaptic Vesicles; Techniques; Testing; Therapeutic; Time; Translating; Translations; United States; Usher Syndrome; Usher Syndrome, Type 1C; Viral Vector; Virus; Visual; Visual impairment; Work; afferent nerve; behavioral response; congenital deafness; deafness; density; early adolescence; early onset; fetal; gene function; gene replacement; gene therapy; genetic manipulation; hearing impairment; in utero; inner ear diseases; intraperitoneal; mRNA Expression; mouse model; neonate; otoconia; postnatal; precursor cell; prenatal therapy; prevent; protein expression; public health relevance; ribbon synapse; round window; seal; sound; spiral ganglion; synaptogenesis; targeted delivery; therapy design

DESCRIPTION (provided by applicant): Bilateral permanent congenital hearing loss is estimated to affect 1-3 neonates per 1,000 live births worldwide. A genetic basis is causal in approximately 50% of the individuals. Treatment options for congenitally deaf or hearing impaired children are limited in scope and efficacy and typically involve hearing aids or cochlear implants. Neither intervention fully restores the richness of native hearing. The conceptual basis of this proposal is to define fetal therapies that correct defects in gene expression or function prior to the onset of overt pathogenic changes in the sensory epithelia of the developing inner ear. We use the mouse as a model system because an ever-expanding array of natural and induced genetic mutations exists that serve as accurate paradigms for human inner ear dysfunction. Our technical approach relies on experimental embryology, a palette of surgical, imaging, microinjection and molecular techniques that permit access to the developing mouse inner ear in utero and enable genetic manipulation of precursor cells that give rise to the auditory and vestibular sensory structures. In this proposal, we aim: 1) to restore auditory function in a mouse model defective in synaptic transmission~ and 2) to restore auditory and vestibular function in a mouse model of Usher syndrome. A missense mutation in the human SLC17A8 gene that encodes vesicular glutamate transporter-3 (VGLUT3) has been associated with an autosomal-dominant form of progressive, high-frequency nonsyndromic deafness. VGLUT3 is selectively localized to inner hair cells in the cochlea and loads the excitatory amino acid neurotransmitter glutamate into synaptic vesicles. The VGLUT3 knockout mouse is born deaf due to the inability of inner hair cells to release glutamate at the afferent nerve terminals. We hypothesize that virus-mediated gene transfer of VGLUT3 to otic epithelial precursors will force VGLUT3 expression in inner hair cells and restore synaptic transmission and hearing in the VGLUT3 knockout mouse. Usher syndrome is the leading genetic cause of combined deafness and blindness and is associated with 13 loci and 10 genes in humans. Three distinct clinical forms are differentiated by the time of onset and severity of auditory, vestibular, and visual dysfunction. Usher syndrome type 1 is the most severe with bilateral profound hearing loss and balance difficulty at birth and retinitis pigmentosa present by early adolescence. The USH1 gene encodes the protein harmonin that is expressed in the sensory hair cell bundle and the ribbon synapse. A single base pair mutation in the harmonin gene creates a cryptic splice site resulting in a frameshift and truncated harmonin protein. The USH1C mouse mutant is born deaf, uncoordinated, and develops vision impairment. We hypothesize that fetal administration of an antisense oligonucleotide will correct harmonin messenger RNA splicing and restore auditory and vestibular function in the USH1C mouse mutant. We are optimistic that the proposed studies will establish the first fetal therapies for congenital deafness and vestibular dysfunction and will help inform translation of these approaches to the clinic.

描述（由申请人提供）：据估计，全世界每1，000例活产中有1-3例新生儿患有双侧永久性先天性听力损失。大约50%的人的遗传基础是因果关系。先天性耳聋或听力受损儿童的治疗方案在范围和疗效方面都很有限，通常涉及助听器或人工耳蜗植入。这两种干预都不能完全恢复原生听觉的丰富性。该建议的概念基础是定义胎儿疗法，在发育中的内耳感觉上皮发生明显的致病性变化之前纠正基因表达或功能的缺陷。我们使用小鼠作为模型系统，因为存在一系列不断扩大的天然和诱导基因突变，这些突变可作为人类内耳功能障碍的准确范例。我们的技术方法依赖于实验胚胎学，手术，成像，显微注射和分子技术的调色板，允许在子宫内进入发育中的小鼠内耳，并实现对产生听觉和前庭感觉结构的前体细胞的遗传操作。在本提案中，我们的目标是：1）恢复突触传递缺陷小鼠模型的听觉功能; 2）恢复Usher综合征小鼠模型的听觉和前庭功能。编码囊泡谷氨酸转运蛋白3（VGLUT 3）的人SLC 17 A8基因的错义突变与常染色体显性形式的进行性、高频非综合征性耳聋相关。VGLUT 3选择性地定位于耳蜗内毛细胞，并将兴奋性氨基酸神经递质谷氨酸加载到突触囊泡中。VGLUT 3基因敲除小鼠天生耳聋，这是由于内毛细胞无法在传入神经末梢释放谷氨酸。 我们假设病毒介导的VGLUT 3基因转移到耳上皮前体细胞将迫使VGLUT 3在内毛细胞中表达，并恢复VGLUT 3敲除小鼠的突触传递和听力。Usher综合征是耳聋和失明的主要遗传原因，与人类的13个位点和10个基因相关。根据听觉、前庭和视觉功能障碍的发作时间和严重程度，可区分三种不同的临床形式。Usher综合征1型是最严重的，出生时双侧严重听力损失和平衡困难，青春期早期出现视网膜色素变性。USH 1基因编码在感觉毛细胞束和带状突触中表达的蛋白harmonin。harmonin基因中的单个碱基对突变产生了一个隐蔽的剪接位点，导致移码和截短的harmonin蛋白。USH 1C突变小鼠天生耳聋，不协调，并出现视力障碍。我们假设，胎儿管理的反义寡核苷酸将纠正harmonin信使RNA剪接和恢复USH 1C小鼠突变的听觉和前庭功能。我们乐观地认为，拟议的研究将建立先天性耳聋和前庭功能障碍的第一个胎儿疗法，并将有助于将这些方法转化为临床。