Pou3f4-expressing Otic Mesenchyme Cells: A Novel Influence for Spiral Ganglion Neuron Survival

表达 Pou3f4 的耳间充质细胞：对螺旋神经节神经元存活的新影响

• 批准号: 10312394
• 负责人: Paige M Brooks
• 金额: $ 3.33万
• 依托单位: GEORGETOWN UNIVERSITY
• 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 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10312394
• 关键词: Adult; Affect; Animals; Apoptosis; Auditory; Auditory Brainstem Responses; Auditory Prosthesis; Auditory system; Award; Biological Assay; Cell physiology; Cells; Clinical; Cochlea; Cochlear Implants; Coculture Techniques; Complement; Cues; Data; Defect; Development; Embryo; Embryonic Development; Environment; Genetic Transcription; Hair Cells; Health; Hearing; Hearing Aids; Human; Image; In Situ Hybridization; In Vitro; Labyrinth; Link; Macrophage Activation; Maryland; Measures; Mediating; Mentors; Mentorship; Mesenchyme; Modeling; Molecular; Morphology; Mus; Mutation; Neurons; Noise; Oxidative Stress; Pattern; Peripheral; Play; Process; Publishing; Reactive Oxygen Species; Research; Research Training; Role; Supporting Cell; Survival Analysis; System; Techniques; Testing; Therapeutic; Time; Tissues; Training; Transcript; Universities; Virus; Work; auditory pathway; axon guidance; base; career; career development; cell type; deafness; experimental study; gene therapy; hearing impairment; improved; in vivo; macrophage; malformation; middle ear; mouse model; mutant; neuron loss; neuronal survival; neurotrophic factor; novel; postnatal; postnatal development; preservation; presynaptic; prevent; spiral ganglion; targeted treatment; transcription factor

PROJECT SUMMARY Hearing function depends on the precise connectivity patterns and proper function of specialized cells in the cochlea including spiral ganglion neurons (SGNs) and their presynaptic partners, hair cells. Deficits in the development, survival or function of these cells underlie hearing impairment and the efficacy of cochlear implants. SGNs in particular form the afferent connection between the peripheral and central auditory systems. SGNs are crucial for proper hearing and are particularly vulnerable to damage. We showed recently that Pou3f4, a transcription factor expressed by otic mesenchyme cells, is normally required for SGN survival during postnatal development. Mutations in Pou3f4 cause human hearing loss, and loss of Pou3f4 in mouse models leads to morphological defects in otic mesenchyme and hearing impairment in addition to decreased neuronal survival. Still, we have a limited understanding of how Pou3f4 regulates these processes because the transcriptional targets of Pou3f4 in the cochlea are not well understood. As described in this proposal, I will first seek to determine the function of Pou3f4 in SGN survival by defining the mechanism of action between otic mesenchyme cells and SGNs. Second, I will investigate the potential for otic mesenchyme cells to promote SGN survival using gene therapy. With respect to the mentored research training aspects of this award, I will receive additional training in virus-mediated gene therapy and auditory brainstem response techniques through co-mentorship by Dr. Ronna Hertzano. We predict that downstream targets of Pou3f4 include trophic or other environmental cues that influence SGN survival. I will take a direct approach to determining these mechanisms using a combination of molecular techniques and live imaging experiments. At the same time, our collaborators will be taking an unbiased, “omics”-based approach to identifying Pou3f4 targets. Additionally, I will reintroduce Pou3f4 and its targets both in vitro and in vivo to determine the extent to which mesenchyme cells promote SGN survival. I will be the one of the first to define how otic mesenchyme cells interact with SGNs to impact neuronal survival in the normally developing cochlea, as well as one of the first to determine the extent to which otic mesenchyme cells could be targeted therapeutically to help promote SGN survival in adults. Thus, this work explores a novel question that will complement ongoing work by others on neurotrophins, gene therapy, cell replacement strategies, and auditory prosthetics. The proposed research, mentoring, and career development aspects of this plan will help prepare me for a successful independent research career.

项目概要 听力功能取决于精确的连接模式和特定细胞的适当功能 耳蜗包括螺旋神经节神经元 (SGN) 及其突触前伙伴毛细胞。赤字 这些细胞的发育、存活或功能是听力障碍和耳蜗功效的基础 植入物。 SGN 特别形成外周听觉系统和中枢听觉系统之间的传入连接。 SGN 对于正常听力至关重要，并且特别容易受到损坏。我们最近表明 Pou3f4 是一种由耳间充质细胞表达的转录因子，通常是 SGN 存活所必需的 在产后发育过程中。 Pou3f4 突变导致人类听力损失，小鼠 Pou3f4 缺失 除了听力下降之外，模型还会导致耳间充质的形态缺陷和听力障碍 神经元存活。尽管如此，我们对 Pou3f4 如何调节这些过程的了解仍然有限，因为 Pou3f4 在耳蜗中的转录靶标尚不清楚。 正如本提案中所述，我将首先通过以下方式寻求确定 Pou3f4 在 SGN 生存中的功能： 定义耳间充质细胞和 SGN 之间的作用机制。其次，我将调查 耳间充质细胞利用基因疗法促进 SGN 存活的潜力。相对于受指导者 在该奖项的研究培训方面，我将接受病毒介导的基因治疗方面的额外培训 由 Ronna Hertzano 博士共同指导的听觉脑干反应技术。我们预测 Pou3f4 的下游目标包括影响 SGN 生存的营养或其他环境因素。我会 采用分子技术和活体技术相结合的直接方法来确定这些机制 成像实验。与此同时，我们的合作者将采取公正的、基于“组学”的方法 识别 Pou3f4 目标。此外，我将重新介绍 Pou3f4 及其体外和体内靶标 确定间充质细胞促进 SGN 存活的程度。我将是第一个定义的人 耳间充质细胞如何与 SGN 相互作用，影响正常发育的耳蜗中的神经元存活， 也是第一个确定耳间充质细胞可以被靶向的程度的人之一 治疗上有助于促进成人 SGN 存活。因此，这项工作探讨了一个新问题 补充其他人在神经营养素、基因治疗、细胞替代策略和听觉方面正在进行的工作 假肢。该计划拟议的研究、指导和职业发展方面将有助于做好准备 我成功的独立研究生涯。