Investigating the Roles of Epithelial Splice Regulatory Proteins in Inner Ear Development

研究上皮剪接调节蛋白在内耳发育中的作用

基本信息

批准号：
9026485
负责人：
ALEX ROHACEK
金额：
$ 4.36万
依托单位：
UNIVERSITY OF PENNSYLVANIA
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-07-01 至 2018-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9026485
关键词：
Address Adult Affect Alternative Splicing Apical Auditory Auditory Brainstem Responses Cadherins Cell Cycle Cell Differentiation process Cell Maturation Cells Cleft Palate Cochlea Cochlear duct Congenital Abnormality Coupled Defect Development Developmental Delay Disorders Duct (organ) structure Embryo Epithelial Epithelium Event Exhibits Exons Family Fibroblast Growth Factor Receptors General Population Genes Goals Hair Cells Head Hearing Hearing Impaired Persons Human Incidence Individual Infant Kinetics Knock-out Knockout Mice Labyrinth Labyrinth Supporting Cells Length Life Link Maintenance Mammals Measures Messenger RNA Morphology Mus Mutant Strains Mice Mutation Myosin ATPase Nature Neonatal Organ Organ of Corti Otic Vesicle Paint Pediatric Hospitals Phase Phenotype Philadelphia Process Protein Isoforms Proteins RNA RNA Splicing RNA-Binding Proteins Role Sensorineural Hearing Loss Sensory Sensory Hair Series Space Perception Specific qualifier value Specificity Structure Supporting Cell Temporal bone structure Testing Time Tissues Transcript Transducers Tube United States Vestibule cell fate specification cell type cranium deafness ear development epithelial to mesenchymal transition exome sequencing gene function genetic regulatory protein hearing impairment improved interest loss of function mutation mutant mutant mouse model next generation sequencing novel precursor cell progenitor public health relevance recombinase research study sound spatiotemporal transcriptome sequencing

项目摘要

DESCRIPTION (provided by applicant): The mammalian inner ear is a dual function organ comprised of the vestibular apparatus that senses spatial orientation and the cochlear duct which functions in sound transduction. The development of the cochlea is of particular interest due to the high incidence of hearing loss that occurs in the general population. The cochlear duct arises from a ventral outgrowth of the inner ear anlage, the otic vesicle. During its development the cochlea extends into a coiled tube that will contain the sensory hair and support cells of the Organ of Corti that are the transducers of sound. The function of the Organ of Corti is tightly coupled to the proper timing of events that occur during its formation. Sensory cell precursors must be specified from the otic epithelium, undergo cell-cycle exit and then differentiate into hair and support cells. The precise timing of these events is crucial for the proper development of the Organ of Corti, the cochlea as a whole, and, subsequently, the sense of hearing. While many of the genes responsible for each of these developmental events have been discovered the factors controlling their timing have yet to be identified. Recently, our collaborator, Dr. Ian Krantz (Children's Hospital of Philadelphia) identified mutations in the Epithelial Splicing Regulatory Protein 1 gene in several individuals with profound sensorineural deafness. In an effort to functionally link the deafness phenotype with mutations in Esrp1 we have begun to analyze mouse mutants in Esrp1 and/or Esrp2 for inner ear phenotypes. My preliminary results indicate that Esrp1 mutants exhibit a truncated cochlear duct and a significant delay in the development of auditory hair and support cells. The goal of this proposal is to determine the mechanism by which Esrp1 regulates the timing of sensory development in the inner ear. To accomplish this objective, I will systematically analyze the inner ears of Esrp mutants for alterations in the specification and cell-cycle exit of sensory progenitors, as well as the differentiation and maturation of cochlear hair and support cells (Aim1a). I will also perform RNAseq experiments on cochlear ducts from Esrp1-/- and control embryos to identify aberrantly spliced mRNA transcripts and potential targets of Esrp1 (Aim 1b). Finally, given the neonatal lethality of Esrp1-/- mutants from a cleft palate defect, I will generate a conditional knockout of Esrp1 in the inner ear to interrogate its requirement for hearing in adult mice (Aim 2). These experiments will uncover unique roles of Esrp genes in inner ear development and highlight the mechanism by which pathogenic mutations in Esrp1 result in deafness.

描述（由应用程序提供）：哺乳动物内耳是一种双重函数，其中包含前庭设备，该设备感受到空间取向和在声音传递中起作用的耳蜗管道。人工耳蜗的发展特别令人感兴趣，这是由于一般人群发生的听力损失发生率很高。耳蜗是由内耳动眼的腹侧生长产生的。在其发育过程中，人工耳蜗延伸到盘绕的管中，该管子将含有感觉的头发和支持Corti器官的细胞，这些细胞是声音的传感器。 Corti器官的功能与形成过程中发生的事件的适当时机紧密耦合。感官细胞前体必须从耳上皮指定，进行细胞周期出口，然后分化为头发和支撑细胞。这些事件的确切时机对于Corti器官的适当发展，整个耳蜗以及随后的听力感至关重要。尽管已经发现了许多负责这些发展事件的基因，但尚未确定控制其时间安排的因素。最近，我们的合作者伊恩·克兰兹（Ian Krantz）博士（费城儿童医院）在几个具有深刻感官死亡的人的上皮剪接调节蛋白1基因中发现了突变。为了在功能上将死亡表型与ESRP1中的突变联系起来，我们已经开始分析ESRP1和/或ESRP2中的小鼠突变体的内耳表型。我的初步结果表明，ESRP1突变体表现出截短的耳蜗管道，并显着延迟听觉头发和支撑细胞。该提案的目的是确定ESRP1调节内耳感觉发展时机的机制。为了实现这一目标，我将系统地分析ESRP突变体的内耳，以改变感觉祖细胞的规范和细胞周期出口，以及耳蜗和支持细胞的分化和成熟（AIM1A）。我还将对来自ESRP1 - / - 和对照胚胎的耳蜗管进行RNASEQ实验，以鉴定ESRP1的异常剪接的mRNA转录本和潜在靶标（AIM 1B）。最后，鉴于ESRP1 - / - 突变体的新生儿致死性来自left裂缺陷，我将产生一个有条件的敲除内耳中的ESRP1询问其在成年小鼠中听力的要求（AIM 2）。这些实验将发现ESRP基因在内耳发育中的独特作用，并强调ESRP1中的致病突变导致耳聋的机制。