Zebrafish sensory hair cell regeneration

斑马鱼感觉毛细胞再生

• 批准号: 9889942
• 负责人: Tatjana Piotrowski
• 金额: $ 35.06万
• 依托单位: STOWERS INSTITUTE FOR MEDICAL RESEARCH
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9889942
• 关键词: Affect; Age; Amphibia; Behavior; Biological Assay; Birds; Bromodeoxyuridine; Cell Death; Cell Differentiation process; Cell Maintenance; Cell Proliferation; Cell physiology; Cells; Complex; Development; Ear; Ensure; Epithelial; Epithelium; Equilibrium; Fishes; Future; Gene Expression; Gene Expression Profiling; Genes; Genetic Epistasis; Genetic Transcription; Goals; Hair Cells; Hearing problem; Heterogeneity; Homeostasis; Human; In Situ Hybridization; In Vitro; Individual; Knowledge; Labyrinth; Life; Location; Maintenance; Mammals; Masks; Modeling; Monitor; Mosaicism; Mus; Mutation; National Institute on Deafness and Other Communication Disorders; Natural regeneration; Pathway interactions; Pharmacologic Substance; Population; Process; Protocols documentation; Regulator Genes; Reporter; Sensory; Sensory Disorders; Sensory Hair; Signal Pathway; Signal Transduction; Skin; Supporting Cell; System; Testing; Therapeutic; Time; Transgenic Organisms; Tretinoin; Vertebrates; WNT Signaling Pathway; Water Movements; Work; Zebrafish; cell type; combinatorial; deaf; deafness; design; disability; embryonic stem cell; experimental study; gene interaction; genetic manipulation; hair cell regeneration; hard of hearing; hearing impairment; human old age (65+); in vivo; induced pluripotent stem cell; inhibitor/antagonist; lateral line; member; mutant; neuromast; notch protein; progenitor; restoration; self-renewal; single cell analysis; stem cells; therapy design; transcriptome; transcriptome sequencing; zebrafish development

Project Summary Hearing loss caused by hair cell degeneration is one of the most widespread sensory disabilities in the world with 30% of people between ages 65 -75 being affected. In humans and mammals, hair cells are unable to regenerate, hence any damage or loss of hair cells is permanent. Therefore, there is an urgent need to develop therapeutic approaches for regenerating hair cells in humans. In contrast to mammals, hair cells in fishes, birds and amphibians regenerate following hair cell death. Very little is known about why hair cells regenerate in some vertebrates and not in others. The goal of this work is to elucidate the gene regulatory network underlying zebrafish hair cell regeneration, The results will help us identify strategies to elicit the restoration of lost hair cells in mammals. Zebrafish possess hair cells in their ears and in their skin as part of the sensory lateral line system that senses water movement. The development of zebrafish lateral line hair cells is controlled by the same signaling pathways as mammalian inner ear hair cells, however, they are easily accessible to direct observation and genetic manipulation. Interactions between Notch and Wnt signaling are crucial for maintaining a balance between progenitor cell self-renewal and differentiation. This work will expand this knowledge by determining how other signaling pathways, such as Fgf and retinoic acid (RA), which also robustly affect hair cell regeneration, fit into this network. The epistatic relationship between the Notch, Wnt, Fgf and RA pathways will be determined by combinatorial manipulation of pathways using mutants, pharmaceutical inhibitors and transgenic lines. In addition, gene interactions that regulate the balance of progenitor cell self-renewal and differentiation will be functionally interrogated at the single cell level in vivo with a combination of time-lapse and cell proliferation/cell fate analyses. An important component of this work is the expression analysis of individual support cells during regeneration and after pathway manipulations. The single cell analysis is powerful, as an unexpected diversity or mosaicism in support cells was discovered, which is masked in bulk support cell RNASeq analyses. In addition, as cells differentiate they will integrate many different signals from other cells over time. These experiments will allow the distinction of cells on different trajectories, temporally order them along those trajectories, and identify new regulatory factors controlling differentiation. The results of these aims will inform us of the gene regulatory network underlying zebrafish hair cell regeneration, which will guide the design of therapies to induce hair cell regeneration in mammals.

项目摘要 毛细胞变性引起的听力损失是世界上最普遍的感觉障碍之一 30％的65-75岁的人受到影响。在人类和哺乳动物中，毛细胞无法 再生，因此毛细胞的任何损害或损失都是永久性的。因此，迫切需要 开发用于再生人毛细胞的治疗方法。与哺乳动物相反，毛细胞中的毛细胞 鱼类，鸟类和两栖动物在毛细胞死亡后再生。关于毛细胞的原因很少 在某些脊椎动物中再生，而不是在其他脊椎动物中再生。这项工作的目的是阐明基因调节 斑马鱼毛细胞再生的网络，结果将有助于我们确定引起的策略 恢复哺乳动物中失去的毛细胞。斑马鱼在耳朵和皮肤中都有毛细胞作为一部分 感觉水运动的感觉侧线系统。斑马鱼侧线头发的发展 细胞由与哺乳动物内耳毛细胞相同的信号通路控制，但是它们很容易 可以直接观察和基因操作。 Notch和Wnt信号之间的相互作用是 对于保持祖细胞自我更新和分化之间的平衡至关重要。这项工作将扩大 通过确定其他信号通路（例如FGF和视黄酸（RA））的知识，这也是如此 牢固地影响毛细胞再生，适合该网络。 Notch，Wnt之间的上任关系 FGF和RA途径将通过使用突变体组合对途径的组合来确定 药物抑制剂和转基因线。另外，调节平衡的基因相互作用 祖细胞的自我更新和分化将在体内在单细胞水平上进行功能询问 延时和细胞增殖/细胞命运分析的组合。这项工作的重要组成部分是 在再生和途径操纵后对单个支持细胞的表达分析。单人 细胞分析是强大的，因为发现了支持细胞中意外的多样性或镶嵌物，这就是 在大量支持细胞RNASEQ分析中掩盖。另外，随着细胞的区分，它们将整合许多 随着时间的推移，其他细胞的不同信号。这些实验将允许细胞在不同的 轨迹，临时沿着这些轨迹订购，并确定控制的新调节因素 分化。这些目标的结果将告知我们斑马鱼的基因调节网络 细胞再生，它将指导疗法设计以诱导哺乳动物的毛细胞再生。