Developing novel stem cell-based epigenetic approaches to treat hearing loss

开发基于干细胞的新型表观遗传学方法来治疗听力损失

• 批准号: 10293594
• 负责人: Zhengqing Hu
• 金额: --
• 依托单位: JOHN D DINGELL VA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-01 至 2023-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10293594
• 关键词: Adult; Affect; Aging; Auditory; Auditory system; Azacitidine; Biological; Brain Stem; Caregivers; Caring; Cell Differentiation process; Cell Line; Cell Lineage; Cells; Clinical; Coculture Techniques; DNA; DNA Methylation; DNA Methylation Regulation; DNA Methyltransferase Inhibitor; DNA Sequence; Data; Deoxycytidine; Department of Defense; Diagnosis; Electrophysiology (science); Epigenetic Process; Epithelial; Epithelial Cells; Exhibits; Family; Functional disorder; Future; Gene Expression; Gene Proteins; Genes; Genomic DNA; Goals; Hair Cells; Health; Hearing; Heritability; Human; In Vitro; Ion Channel; Labyrinth; Medical; Medical center; Methylation; Military Personnel; Mus; Natural regeneration; Neurons; Noise; Pattern; Phenotype; Pilot Projects; Potassium; Proteins; Quality of life; RNA; Reporting; Role; Safety; Science; Sensory Hair; Signal Transduction; Stem Cell Research; Supporting Cell; Synapses; System; Testing; Tinnitus; Transplantation; Traumatic Brain Injury; United States Department of Veterans Affairs; United States Food and Drug Administration; Utricle structure; Veterans; Work; base; clinical application; deaf; demethylation; experience; hearing impairment; hearing restoration; improved; in vivo; inhibitor; inner ear development; inner ear diseases; innovation; mechanotransduction; military service; military veteran; novel; novel strategies; permanent hearing loss; receptor; service member; sound; spiral ganglion; stem; stem cell based approach; stem cell technology; stem cells; success; synaptogenesis; welfare

Project Summary Auditory hair cells are mechanotransduction receptors that convey sound signals to the brainstem via spiral ganglion neurons (SGNs). However, hair cells are vulnerable to a number of insults, including noise, blast, aging, and traumatic brain injury. Adult mammalian hair cell loss is usually irreversible, causing permanent hearing loss and other inner ear disorders. Currently, there are no biological approaches to replenish human hair cells. The long-term goal of this project is to use stem cell-based approaches to regenerate the damaged auditory system to treat hearing loss and other inner ear disorders. In this proposal, we will generate human sensory hair cells and rebuild hair cell-SGN connections using a stem cell-based epigenetic approach. We have identified human utricle epithelia-derived prosensory-like cells (HUCs) that exhibit features of prosensory cells. We found that HUCs demonstrated genomic DNA methylation in a pilot study. We applied the DNA methyltransferase (DNMT) inhibitor 5-azacytidine (5-aza) to HUCs and found that treated HUCs expressed hair cell genes and proteins. Therefore, in this proposal we hypothesize that the DNMT inhibitor 5- aza induces HUCs to differentiate into new functional hair cells that can replace damaged hair cells and form synapses with auditory neurons. To test this hypothesis, three specific aims are proposed: Specific Aim 1: Define the induction of HUCs into sensory hair cells by 5-aza. Specific Aim 2: Investigate the integration and synapse formation of 5-aza-treated HUCs in vitro. Specific Aim 3: Determine the survival, differentiation, integration, and function of 5-aza-treated HUCs in vivo. In this proposal, we will generate human sensory hair cells and rebuild hair cell-neuron connections using a stem cell-based epigenetic approach. The advantage of an epigenetic approach is that expression and phenotype of gene are heritably changed, whereas their DNA sequences remain constant. The long-term aims of this work are (a) to discover and explore innovative regeneration approaches to improve the health and welfare of military personnel and veterans with hearing disturbances, and (b) to accelerate the transition of stem cell technologies into new standards of care to treat hearing loss and other inner ear disorders. Success of this work will significantly promote the research of stem cell-based hearing regeneration, which will potentially affect the state of medical science in today’s battlefield experience on function, wellness, and overall quality of life for veterans as well as their caregivers and families.

项目摘要 听觉毛细胞是机械转导的受体，可通过螺旋传递声音信号到脑干 神经神经元（SGNS）。但是，毛细胞容易受到许多伤害，包括噪声，爆炸， 衰老和创伤性脑损伤。成人哺乳动物毛细胞损失通常是不可逆的，导致永久性 听力损失和其他内耳障碍。目前，没有复制人类的生物学方法 毛细胞。该项目的长期目标是使用基于干细胞的方法来再生受损的 听觉系统治疗听力损失和其他内耳障碍。在此提案中，我们将产生人类 感觉毛细胞并使用基于干细胞的表观遗传学方法来重建毛细胞-SGN连接。 我们已经确定了人类肠上皮衍生的类带状细胞（HUC），暴露了 冒充细胞。我们发现，在一项初步研究中，HUCS表现出基因组DNA甲基化。我们应用了 DNA甲基转移酶（DNMT）抑制剂5-氮杂丁胺（5-aza）到HUCS，发现已处理过的HUCS 表达的毛细胞基因和蛋白质。因此，在此提案中，我们假设DNMT抑制剂5-- AZA影响HUCS以区分新功能性毛细胞，这些毛细胞可以替代受损的毛细胞并形成 与听觉神经元的突触。为了检验这一假设，提出了三个具体目标： 特定目标1：通过5-Za定义HUC对感觉毛细胞的诱导。 具体目标2：研究体外5-Aza处理的HUC的整合和突触形成。 特定目标3：确定经体内5-aza处理的HUC的生存，分化，整合和功能。 在此提案中，我们将生成人类的感觉毛细胞，并使用 基于干细胞的表观遗传学方法。表观遗传方法的优点是表达和 基因的表型可遗传，而其DNA序列保持恒定。长期目标 这项工作是（a）发现和探索创新的再生方法，以改善健康和 有听力灾难的军事人员和退伍军人的福利，（b）加速过渡 干细胞技术成为新的护理标准，以治疗听力损失和其他内耳障碍。成功 这项工作将大大促进基于干细胞的听力再生的研究，这将 在当今战场上有关功能，健康和整体的战场经验中，有可能影响医学科学的状态 退伍军人以及他们的照顾者和家庭的生活质量。