Foxo3 Mechanisms in Noise Damage

Foxo3 噪声损害机制

• 批准号: 10640125
• 负责人: Patricia M. White
• 金额: $ 31.48万
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640125
• 关键词: Ablation; Actin-Binding Protein; Actins; Address; Adult; Affect; Alleles; American; Animal Model; Apoptosis; Autophagocytosis; Binding; Biological Assay; Biological Markers; Breeding; CRISPR/Cas technology; Caspase; Cell Death; Cell Survival; Cells; Cessation of life; Clustered Regularly Interspaced Short Palindromic Repeats; Cochlea; DNA; Data; Disabled Persons; Distress; Enterobacteria phage P1 Cre recombinase; FOXO3A gene; Frequencies; Genes; Genetic; Genetic Polymorphism; Genetic Variation; Goals; Grant; Hearing; Hearing Protection; Hearing Tests; Heat-Shock Proteins 70; Homeostasis; Hour; Human; Human Genetics; Individual; Induction of Apoptosis; Inner Hair Cells; Knock-out; Knockout Mice; Knowledge; Link; LoxP-flanked allele; Measures; Methods; Modeling; Modification; Mus; Neurons; Noise; Noise-Induced Hearing Loss; Nuclear; Nucleic Acid Regulatory Sequences; Occupational; Occupational Noise; Ontology; Outer Hair Cells; Oxidative Stress; Persons; Population; Predisposition; Proteins; RNA; RNA analysis; Recovery; RiboTag; Ribosomes; Risk; Role; Single Nucleotide Polymorphism; Stimulus; Supporting Cell; Technology; Testing; Validation; Variant; Wild Type Mouse; Work; Workplace; biological adaptation to stress; biomarker identification; cell type; conditional knockout; deaf; disability; experimental study; gain of function; genetic variant; hearing impairment; hearing preservation; loss of function; mouse model; mutant; noise exposure; novel; overexpression; preservation; programs; protein expression; rat Pres protein; recombinase; response; sound; spiral ganglion; stress reduction; transcription factor; transcriptome sequencing; translatome

Noise induced hearing loss (NIHL) has disabled millions of people world-wide. Individual risk for NIHL varies from person to person under similar exposure conditions, suggesting that genetic factors contribute to susceptibility. We have found that mice lacking a transcription factor called FOXO3 become severely and permanently deafened after a noise exposure that only briefly affects their wild-type littermates. FOXO3 has multiple functions in other cell types, including oxidative stress reduction, autophagy, and directly inducing apoptosis. A recent study linked human genetic variations in FOXO3 to a greater susceptibility to occupational NIHL. However, the FOXO3 alleles associated with NIHL drive increased expression of FOXO3. Thus, there is evidence to indicate that FOXO3 is important for hearing preservation, but there is also evidence that excess FOXO3 drives NIHL. In this grant, we seek to address this knowledge gap by researching the mechanisms of FOXO3 function, using translatome sequencing, cell-specific Foxo3 conditional knockouts (cKO), and CRISPR modifications to generate mouse lines that can be used to investigate the human NIHL-linked FOXO3 allele. In Foxo3-knockout (KO) mice, noise eliminates high-frequency outer hair cells (OHCs). We show that this occurs through a rapid cell death program called parthanatos, which is caspase-independent apoptosis. Parthanatos indicates that in the absence of any noise damage, Foxo3-KO OHCs are primed for death. Bulk RNA-Seq data from control Foxo3-KO and wild-type littermates show no evidence for changes in oxidative stress reducers known to be regulated by FOXO3. Instead, we see changes in actin binding genes expressed in OHCs. In Aim 1, we propose to validate this screen and identify markers of OHC distress in the Foxo3-KO through translatome sequencing. In Aim 2, we propose to make cell-specific Foxo3-cKO to identify the cells in which FOXO3 acts. Wild-type mice express FOXO3 protein in both OHCs and in the surrounding supporting cells (SCs). By using inducible DNA recombinases lines specific to either OHCs or SCs, we can ablate FOXO3 function in either cell type. We will expose such Foxo3-cKO mice to noise and determine their NIHL susceptibility. Finally, in Aim 3, we have used CRISPR genetic modification technology to create two mouse lines, one with control sequences (Foxo3-T-allele mice), as well as one homologous to the human FOXO3 allele that confers NIHL susceptibility (Foxo3-G-allele mice). We will validate that the Foxo3-G-allele mouse line has increased levels of FOXO3 in cochlear cells after noise exposure. We hypothesize that this modification promotes apoptosis from FOXO3 activation, and we will test that hypothesis by exposing Foxo3- G-allele mice to noise, measuring their hearing and analyzing potential cellular losses. In sum, through both loss-of-function and gain-of-function experiments, we will analyze FOXO3's role in hearing loss from noise.

噪声性听力损失（NIHL）已使全世界数百万人致残。NIHL的个体风险

项目成果

Semi-Automated Analysis of Peak Amplitude and Latency for Auditory Brainstem Response Waveforms Using R.

使用R的听觉脑干响应波形的峰值振幅和潜伏期的半自动分析。

• DOI: 10.3791/64737
• 发表时间: 2022-12-09
• 期刊: Journal of visualized experiments : JoVE
• 作者: Na D;White PM
• 通讯作者: White PM

Single cell RNA sequencing analysis of mouse cochlear supporting cell transcriptomes with activated ERBB2 receptor indicates a cell-specific response that promotes CD44 activation.

• DOI: 10.3389/fncel.2022.1096872
• 发表时间: 2022
• 期刊: FRONTIERS IN CELLULAR NEUROSCIENCE
• 影响因子: 5.3
• 作者: Piekna-Przybylska, Dorota;Na, Daxiang;Zhang, Jingyuan;Baker, Cameron;Ashton, John M.;White, Patricia M.
• 通讯作者: White, Patricia M.