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受体，通过螺旋将声音信号传递到脑干 神经节神经元(SGN)。然而，毛细胞很容易受到许多侮辱，包括噪音、爆炸、 衰老和创伤性脑损伤。成年哺乳动物的毛细胞丢失通常是不可逆转的，导致永久性的 听力损失和其他内耳疾病。目前，还没有生物方法来补充人类 毛细胞。这个项目的长期目标是使用基于干细胞的方法来再生受损的 用于治疗听力损失和其他内耳疾病的听觉系统。在这项提案中，我们将产生人类 感觉毛细胞和重建毛细胞-SGN连接使用基于干细胞的表观遗传学方法。 我们已经鉴定出人类椭圆形上皮来源的感觉样细胞(HUCS)，它具有以下特征 前感官细胞。我们发现，在一项初步研究中，HUCS显示了基因组DNA的甲基化。我们应用了 DNA甲基转移酶(DNMT)抑制剂5-氮杂胞苷(5-aza)对HUCS的作用 表达毛细胞基因和蛋白质。因此，在这项提议中，我们假设DNMT抑制剂5- Aza诱导HUCS分化为新的功能毛细胞，可以取代受损的毛细胞并形成 与听觉神经元的突触。为了验证这一假设，我们提出了三个具体目标： 具体目标1：明确5-氮杂氮对HUCS向感觉毛细胞的诱导作用。 特定目的2：研究5-氮杂氮处理的HUC在体外的整合和突触形成。 具体目标3：确定5-氮杂氮处理的HUC在体内的存活、分化、整合和功能。 在这项计划中，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。表观遗传学方法的优点是表达和 基因的表型是遗传改变的，而它们的DNA序列保持不变。长期目标 这项工作的主要内容是：(A)发现和探索创新的再生方法，以改善健康和 为有听力障碍的军事人员和退伍军人提供福利，以及(B)加快过渡 干细胞技术成为治疗听力损失和其他内耳疾病的新标准。成功 这项工作将极大地促进基于干细胞的听力再生的研究，这将 潜在地影响当今战场上的医学状态在功能、健康和整体上的体验 退伍军人及其照顾者和家人的生活质量。