Understanding the mechanism of Sox2 haploinsufficiency on supporting cell to hair cell conversion in the mature mouse cochlea.

了解 Sox2 单倍体不足支持成熟小鼠耳蜗细胞向毛细胞转化的机制。

• 批准号: 10189501
• 负责人: Melissa McGovern
• 金额: $ 6.91万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189501
• 关键词: ATAC-seq; Adopted; Affect; Aging; Alleles; Animals; Binding; Binding Sites; Cells; Cessation of life; ChIP-seq; Chickens; Chromatin; Cochlea; Data; Development; Differentiated Gene; Diphtheria Toxin; Dose; GFI1 gene; Gene Expression; Genes; Genetic; Genetically Engineered Mouse; Goals; Hair Cells; Inner Hair Cells; Knock-in Mouse; Labyrinth; Life; Loudness; Maps; Modeling; Molecular Genetics; Mus; Natural regeneration; Neonatal; Noise; Occupational Noise; Organ of Corti; Pharmaceutical Preparations; Phenotype; Process; Production; RNA; Regenerative research; Regulation; Reporter; Research; Role; Sampling; Sensory Hair; Signal Transduction; Source; Supporting Cell; Testing; Therapeutic; Tissues; Vertebrates; XCL1 gene; Zebrafish; cell killing; epigenetic regulation; experimental study; gene therapy; genomic locus; hair cell regeneration; hearing impairment; hearing loss treatment; hearing restoration; neonatal mice; ototoxicity; overexpression; permanent hearing loss; prevent; transcription factor; transcriptome; transcriptome sequencing

Abstract Hearing loss affects millions of people worldwide and can result from ototoxic medications, recreational or occupational noise exposure, as well as aging; currently, there is no treatment for hearing loss. Recent advances suggest that efforts to treat hearing loss may benefit from a gene therapy approach. However, strategies to promote the regeneration of sensory hair cells in the mature cochlea have yet to provide a reliable therapy for hearing loss. Research initially focused on the hair cell inducing transcription factor Atoh1, which rapidly converts neighboring supporting cells into hair cells in neonatal tissue but has produced limited results in the mature cochlea. More recent evidence supports a multifactor approach that involves the modulation of both hair cell and supporting cell genes within supporting cells. This approach is still limited, as it does not produce cells which resemble normal hair cells. Previous evidence indicated that the loss of one allele of Sox2 results in a haploinsufficient phenotype that produced extra inner hair cells during development as well as enhancing regeneration observed in the neonatal mouse cochlea. This study will extend these findings by using genetically engineered mouse lines to investigate the mechanism of Sox2 haploinsufficiency in the mature cochlea both functionally and genetically. My preliminary data suggests that Sox2 haploinsufficiency promotes the reprogramming of supporting cells into hair cells in the undamaged cochlea after the expression of the hair cell transcription factors Atoh1, Gfi1, and Pou4f3. Aim 1 of this study will investigate whether Sox2 haploinsufficiency can prime supporting cells to respond to these hair cell transcription factors and regenerate lost hair cells. Aim 2 of this study will explore the mechanism by which Sox2 haploinsufficiency promotes the conversion of supporting cells into hair cells by assessing both direct and indirect targets of Sox2 as well as the overall changes in gene expression profiles. To do this, I will perform RNA, ATAC, and CUT & RUN-ChIP- sequencing on purified mature supporting cells to assess the consequences of Sox2 haploinsufficiency.

摘要 听力损失影响着全世界数百万人，可能是由耳毒性药物、娱乐性药物、 或职业噪声暴露，以及老化;目前，没有治疗听力损失的方法。最近 研究进展表明，基因疗法可能会对治疗听力损失有所帮助。然而，在这方面， 促进成熟耳蜗中感觉毛细胞再生的策略尚未提供可靠的 听力损失的治疗研究最初集中在毛细胞诱导转录因子Atoh 1， 在新生儿组织中迅速将邻近的支持细胞转化为毛细胞，但效果有限 在成熟的耳蜗中。最近的证据支持一种多因素的方法，涉及调节 支持细胞内的毛细胞和支持细胞基因。这种方法仍然是有限的，因为它不 产生类似于正常毛细胞的细胞。以前的证据表明，Sox 2基因的一个等位基因的缺失， 导致在发育过程中产生额外的内毛细胞的单倍不足表型， 增强新生小鼠耳蜗中观察到的再生。这项研究将通过以下方式扩展这些发现： 使用基因工程小鼠系研究Sox 2单倍不足的机制， 成熟的耳蜗在功能上和基因上。我的初步数据表明Sox 2单倍不足 促进表达后未受损耳蜗中支持细胞重编程为毛细胞 毛细胞转录因子Atoh 1、Gfi 1和Pou 4f 3。本研究的目的1将研究Sox 2是否 单倍型不足可以引发支持细胞对这些毛细胞转录因子做出反应并再生 失去了毛细胞本研究的目的2将探讨Sox 2单倍不足促进细胞凋亡的机制。 通过评估Sox 2的直接和间接靶点，以及 基因表达谱的总体变化。要做到这一点，我将执行RNA，ATAC，切割和运行ChIP- 对纯化的成熟支持细胞进行测序以评估Sox 2单倍不足的后果。